Pharmaceutical compositions of therapeutically active compounds

ABSTRACT

Provided are compounds and pharmaceutical compositions useful for treating cancer and methods of treating cancer comprising administering to a subject in need thereof a compound or pharmaceutical composition described herein.

CLAIM OF PRIORITY

This application is a continuation of U.S. Ser. No. 16/460,111, filedJul. 2, 2019, which is a continuation of U.S. Ser. No. 15/949,750 filedApr. 10, 2018, which is a continuation of U.S. Ser. No. 15/125,880,filed Sep. 13, 2016, which is a national stage application under 35U.S.C. § 371 of International Application No. PCT/US2015/020349, filedMar. 13, 2015, which claims priority from U.S. Ser. No. 61/953,487 filedMar. 14, 2104 and U.S. Ser. No. 62/081,542 filed Nov. 18, 2014, each ofwhich is incorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION

Isocitrate dehydrogenases (IDHs) catalyze the oxidative decarboxylationof isocitrate to 2-oxoglutarate (i.e., α-ketoglutarate). These enzymesbelong to two distinct subclasses, one of which utilizes NAD(+) as theelectron acceptor and the other NADP(+). Five isocitrate dehydrogenaseshave been reported: three NAD(+)-dependent isocitrate dehydrogenases,which localize to the mitochondrial matrix, and two NADP(+)-dependentisocitrate dehydrogenases, one of which is mitochondrial and the otherpredominantly cytosolic. Each NADP(+)-dependent isozyme is a homodimer.

IDH1 (isocitrate dehydrogenase 1 (NADP+), cytosolic) is also known asIDH; IDP; IDCD; IDPC or PICD. The protein encoded by this gene is theNADP(+)-dependent isocitrate dehydrogenase found in the cytoplasm andperoxisomes. It contains the PTS-1 peroxisomal targeting signalsequence. The presence of this enzyme in peroxisomes suggests roles inthe regeneration of NADPH for intraperoxisomal reductions, such as theconversion of 2, 4-dienoyl-CoAs to 3-enoyl-CoAs, as well as inperoxisomal reactions that consume 2-oxoglutarate, namely thealpha-hydroxylation of phytanic acid. The cytoplasmic enzyme serves asignificant role in cytoplasmic NADPH production.

The human IDH1 gene encodes a protein of 414 amino acids. The nucleotideand amino acid sequences for human IDH1 can be found as GenBank entriesNM_005896.2 and NP_005887.2 respectively. The nucleotide and amino acidsequences for IDH1 are also described in, e.g., Nekrutenko et al., Mol.Biol. Evol. 15:1674-1684(1998); Geisbrecht et al., J. Biol. Chem.274:30527-30533(1999); Wiemann et al., Genome Res. 11:422-435(2001); TheMGC Project Team, Genome Res. 14:2121-2127(2004); Lubec et al.,Submitted (December 2008) to UniProtKB; Kullmann et al., Submitted (June1996) to the EMBL/GenBank/DDBJ databases; and Sjoeblom et al., Science314:268-274(2006).

Non-mutant, e.g., wild type, IDH1 catalyzes the oxidativedecarboxylation of isocitrate to α-ketoglutarate thereby reducingNAD+(NADP+) to NADH (NADPH), e.g., in the forward reaction:

Isocitrate+NAD⁺(NADP+)→α-KG+CO₂+NADH(NADPH)+H⁺.

It has been discovered that mutations of IDH1 present in certain cancercells result in a new ability of the enzyme to catalyze theNAPH-dependent reduction of α-ketoglutarate to R(−)-2-hydroxyglutarate(2HG). The production of 2HG is believed to contribute to the formationand progression of cancer (Dang, L et al, Nature 2009, 462:739-44).

The inhibition of mutant IDH1 and its neoactivity is therefore apotential therapeutic treatment for cancer. Accordingly, there is anongoing need for inhibitors of IDH1 mutants having alpha hydroxylneoactivity.

PCT Publication No. WO 2013/107291 and US Publication No. US2013/0190249 hereby incorporated by reference in their entirety,disclose compounds that inhibit IDH1 mutants (e.g., IDH1R132H orIDH1R132C). These applications additionally disclose methods for thepreparation of inhibitors of mutant IDH1, pharmaceutical compositionscontaining these compounds, and methods for the therapy of diseases,disorders, or conditions (e.g., cancer) associated with overexpressionand/or amplification of mutant IDH1.

There is a need for pharmaceutical compositions that would haveproperties suitable for large-scale manufacturing and formulation, aswell as utility in treating advanced hematologic malignancies, such asacute myelogenous leukemia (AML), myelodysplastic syndrome (MDS),myeloproliferative neoplasms (MPN), myeloproliferative neoplasms (MPN),chronic myelomonocytic leukemia (CMML), B-acute lymphoblastic leukemias(B-ALL), or lymphoma (e.g., T-cell lymphoma), each characterized by thepresence of a mutant allele of IDH1.

SUMMARY OF INVENTION

Disclosed herein are methods of treating advanced hematologicmalignancies, such as acute myelogenous leukemia (AML), myelodysplasticsyndrome (MDS), myeloproliferative neoplasms (MPN), myeloproliferativeneoplasms (MPN), chronic myelomonocytic leukemia (CMML), B-acutelymphoblastic leukemias (B-ALL), or lymphoma (e.g., T-cell lymphoma),each characterized by the presence of a mutant allele of IDH1,comprising, administering to a subject in need thereof a soliddispersion or a pharmaceutical composition comprising a soliddispersion, and at least one pharmaceutically acceptable carrier. Insome embodiments, the advanced hematologic malignancies arecharacterized by a mutant allele of IDH1, wherein the IDH1 mutationresults in a new ability of the enzyme to catalyze the NAPH-dependentreduction of α-ketoglutarate to R(−)-2-hydroxyglutarate (2HG) in apatient. In one embodiment, the mutant IDH1 has an R132X mutation. Inone embodiment, the R132X mutation is selected from R132H, R132C, R132L,R132V, R132S and R132G. In one embodiment, the R132X mutation is R132Hor R132C. In one embodiment, the R132X mutation is R132H. In someembodiments, the advanced hematologic malignancies harbor a co-mutation,e.g., a co-mutation selected from NPM1, FLT3, TET2, CEBPA, DNMT3A, andMLL.

In one aspect, the present invention provides a method of evaluating asubject, the method comprising: acquiring, e.g., directly acquiring, avalue for the level of a compound(S)-N-((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide(Compound 1), or a pharmaceutically acceptable salt thereof; or thelevel of an alpha hydroxy neoactivity product, e.g., 2HG, e.g., R-2HG(2HG), in the subject, that has been treated with Compound 1, to therebyevaluate the subject.

In another aspect, the present invention provides a method of evaluatinga subject, the method comprising: administering to the subject in needthereof a compound(S)-N-((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide(Compound 1), or a pharmaceutically acceptable salt thereof, andacquiring a value for the level of Compound 1 or the level of an alphahydroxy neoactivity product, e.g., 2HG, e.g., R-2HG (2HG), in thesubject, to thereby evaluate the subject.

In some embodiments, acquiring comprises receiving a sample from thesubject. In some embodiments, acquiring comprises transmitting the valueto another party, e.g., the party that administered Compound 1.

In some embodiments, the value for the level of Compound 1 is acquiredby analyzing the concentration of Compound 1 in a bodily fluid, e.g.,blood, plasma or urine. In some embodiments, the value for the level ofCompound 1 is acquired by analyzing the level of Compound 1 in bonemarrow, e.g., analyzing a sample from a bone marrow biopsy and/oraspirate for the level of Compound 1.

In some embodiments, the value for the level of 2HG is acquired byanalyzing the concentration of 2HG in a bodily fluid, e.g., blood,plasma or urine. In some embodiments, the value for the level of 2HG isacquired by analyzing the level of 2HG in bone marrow, e.g., analyzing asample from a bone marrow biopsy and/or aspirate for the level of 2HG.

In some embodiments, the analysis is performed by sample analysis ofbodily fluid, such as blood, plasma or urine, by e.g., a chromatographicmethod, e.g., mass spectroscopy, e.g. LC-MS. In some embodiments, theanalysis is performed by spectroscopic analysis, e.g., magneticresonance-based analysis, e.g., MRI and/or MRS measurement.

In some embodiments, the subject has been administered Compound 1 lessthan about 30 days prior to the evaluation, e.g., less than about 29days, e.g., less than about 28 days, e.g., less than about 27 days,e.g., less than about 26 days, e.g., less than about 25 days, less thanabout 24 days, e.g., less than about 23 days, e.g., less than about 22days, e.g., less than about 21 days, e.g., less than about 20 days,e.g., less than about 19 days, e.g., less than about 18 days, e.g., lessthan about 17 days, e.g., less than about 16 days, e.g., less than about15 days, e.g., less than about 14 days, e.g., about 7 days, less thanabout 6 days, less than about 5 days, less than about 4 days, less thanabout 3 days, or less than 72 hours prior to the evaluation, e.g., lessthan 48 hours, less than 24 hours, less than 12 hours, less than 10hours, less than 8 hours, less than 6 hours, less than 4 hours, lessthan 3 hours, less than 2 hours, less than 1.5 hours, less than 1 hour,less than 45 minutes, less than 30 minutes, or less than 15 minutes,prior to the evaluation.

In some embodiments, the subject has been administered, e.g., orally,Compound 1 at a dose of about 10 mg to about 3000 mg, e.g., once ortwice daily, (e.g., about every 8-16 hours, e.g., about every 12 hours),or (e.g., about every 12-36 hours, e.g., about every 24 hours), e.g., atabout 10 mg to about 60 mg, at about 60 mg to about 200 mg, at about 200mg to about 500 mg, at about 500 mg to about 1200 mg, at about 1200 mgto about 2000 mg, or at about 2000 mg to about 3000 mg, e.g., at about50 mg, at about 100 mg, at about 300 mg, at about 500 mg, at about 800mg once or twice daily, e.g., about every 12 hours, or e.g., about every24 hours, prior to the evaluation.

In some embodiments, the subject has or is diagnosed as having adisorder. In some embodiments, the disorder is an advanced hematologicmalignancy, e.g., an advanced hematologic malignancy characterized bythe presence of a mutant allele of IDH1. In some embodiments, theadvanced hematologic malignancy is characterized by a mutant allele ofIDH1, wherein the IDH1 mutation results in a new ability of the enzymeto catalyze the NAPH-dependent reduction of α-ketoglutarate toR(−)-2-hydroxyglutarate (2HG) in a patient. In one embodiment, themutant IDH1 has an R132X mutation. In one embodiment, the R132X mutationis selected from R132H, R132C, R132L, R132V, R132S and R132G. In oneembodiment, the R132X mutation is R132H or R132C. In one embodiment, theR132X mutation is R132H. In some embodiments, the advanced hematologicmalignancy harbors a co-mutation, e.g., a co-mutation selected fromNPM1, FLT3, TET2, CEBPA, DNMT3A, and MLL.

In some embodiments, the disorder is selected from acute myelogenousleukemia (AML), myelodysplastic syndrome (MDS), myeloproliferativeneoplasms (MPN), myeloproliferative neoplasms (MPN), chronicmyelomonocytic leukemia (CMML), B-acute lymphoblastic leukemias (B-ALL),B-acute lymphoblastic leukemias (B-ALL), and lymphoma (e.g., T-celllymphoma), wherein each is characterized by the presence of a mutantallele of IDH1. In some embodiments, the disorder is selected fromadvanced IDH1 mutation-positive relapsed and/or refractory AML (R/RAML), untreated AML, and MDS.

In some embodiments, the subject has been previously treated with one ormore chemotherapeutic agent(s). In some embodiments, thechemotherapeutic agent is selected from cytarabine (Ara-C),daunorubicin, etoposide, mitoxantrone, idarubicin, 5-azacytidine,decitabine, SGN33A, sargramostim, WT-1 analog peptide vaccine,tipifarnib, MK-8242, campath, and 6 Mercaptopurine (6MP).

In another aspect, the present invention provides a method of evaluatinga subject, the method comprising: acquiring, e.g., directly acquiring, avalue for the level of blast cells, e.g., leukemic blast cells, e.g.,myeloblasts or myeloid blasts, in the subject, that has been treatedwith a compound(S)-N-((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide(Compound 1), to thereby evaluate the subject.

In another aspect, the present invention provides a method of evaluatinga subject, the method comprising: administering to the subject in needthereof a compound(S)-N-((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide(Compound 1), or a pharmaceutically acceptable salt thereof, andacquiring a value for the level of blast cells, e.g., leukemic blastcells, e.g., myeloblasts or myeloid blasts, in the subject, to therebyevaluate the subject.

In some embodiments, acquiring comprises receiving a sample from thesubject. In some embodiments, acquiring comprises transmitting the valueto another party, e.g., the party that administered Compound 1.

In some embodiments, the evaluation comprises acquiring a value for thelevel of blast cells, e.g., leukemic blast cells, e.g., myeloblasts ormyeloid blasts, e.g., a blast cell count, in a sample from the subject,and comparing the value to a reference standard. In some embodiments,the reference standard is the total number of cells in the sample. Insome embodiments, the sample comprises blast cells, myelocytes,neutrophils, promyelocytes, metamyelocytes, and monocytes.

In some embodiments, the value for the level of blast cells, e.g.,leukemic blast cells, e.g., myeloblasts or myeloid blasts, is acquiredby analyzing the bone marrow, e.g., by analyzing blast counts in bonemarrow aspirates. In some embodiments, the bone marrow is analyzed,e.g., about every two weeks, e.g., (between days 12-18, e.g, on day 15),(between days 26-32, e.g., on day 29), (between days 54-60, e.g., on day57), and then about every 50-60 days thereafter, e.g., every 56 daysthereafter, e.g., on days 15, 29 and 57, and then every 56 daysthereafter.

In some embodiments, the subject has been administered Compound 1 lessthan about 30 days prior to the evaluation, e.g., less than about 29days, e.g., less than about 28 days, e.g., less than about 27 days,e.g., less than about 26 days, e.g., less than about 25 days, less thanabout 24 days, e.g., less than about 23 days, e.g., less than about 22days, e.g., less than about 21 days, e.g., less than about 20 days,e.g., less than about 19 days, e.g., less than about 18 days, e.g., lessthan about 17 days, e.g., less than about 16 days, e.g., less than about15 days, e.g., less than about 14 days, e.g., about 7 days, less thanabout 6 days, less than about 5 days, less than about 4 days, less thanabout 3 days, or less than 72 hours prior to the evaluation, e.g., lessthan 48 hours, less than 24 hours, less than 12 hours, less than 10hours, less than 8 hours, less than 6 hours, less than 4 hours, lessthan 3 hours, less than 2 hours, less than 1.5 hours, less than 1 hour,less than 45 minutes, less than 30 minutes, or less than 15 minutes,prior to the evaluation.

In some embodiments, the subject has been administered, e.g., orally,Compound 1 at a dose of about 10 mg to about 3000 mg, e.g., once ortwice daily, (e.g., about every 8-16 hours, e.g., about every 12 hours),or (e.g., about every 12-36 hours, e.g., about every 24 hours), e.g., atabout 10 mg to about 60 mg, at about 60 mg to about 200 mg, at about 200mg to about 500 mg, at about 500 mg to about 1200 mg, at about 1200 mgto about 2000 mg, or at about 2000 mg to about 3000 mg, e.g., at about50 mg, at about 100 mg, at about 300 mg, at about 500 mg, at about 800mg once or twice daily, e.g., about every 12 hours, or e.g., about every24 hours, prior to the evaluation.

In some embodiments, the subject has or is diagnosed as having adisorder. In some embodiments, the disorder is an advanced hematologicmalignancy, e.g., an advanced hematologic malignancy characterized bythe presence of a mutant allele of IDH1. In some embodiments, theadvanced hematologic malignancy is characterized by a mutant allele ofIDH1, wherein the IDH1 mutation results in a new ability of the enzymeto catalyze the NAPH-dependent reduction of α-ketoglutarate toR(−)-2-hydroxyglutarate (2HG) in a patient. In one embodiment, themutant IDH1 has an R132X mutation. In one embodiment, the R132X mutationis selected from R132H, R132C, R132L, R132V, R132S and R132G. In anotheraspect, the R132X mutation is R132H or R132C. In one embodiment, theR132X mutation is R132H.

In some embodiments, the advanced hematologic malignancy ischaracterized by a co-mutation, e.g., a co-mutation selected from NPM1,FLT3, TET2, CEBPA, DNMT3A, and MLL.

In some embodiments, the disorder is selected from acute myelogenousleukemia (AML), myelodysplastic syndrome (MDS), myeloproliferativeneoplasms (MPN), myeloproliferative neoplasms (MPN), chronicmyelomonocytic leukemia (CMML), B-acute lymphoblastic leukemias (B-ALL),B-acute lymphoblastic leukemias (B-ALL), and lymphoma (e.g., T-celllymphoma), wherein each is characterized by the presence of a mutantallele of IDH1. In some embodiments, the disorder is selected fromadvanced IDH1 mutation-positive relapsed and/or refractory AML (R/RAML), untreated AML, and MDS.

In some embodiments, the subject has been previously treated with one ormore chemotherapeutic agent(s). In some embodiments, thechemotherapeutic agent is selected from cytarabine (Ara-C),daunorubicin, etoposide, mitoxantrone, idarubicin, 5-azacytidine,decitabine, SGN33A, sargramostim, WT-1 analog peptide vaccine,tipifarnib, MK-8242, campath, and 6 Mercaptopurine (6MP).

In another aspect, the present invention provides a method of treating adisorder in a subject, the method comprising: administering to thesubject in need thereof a compound(S)-N-((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide(Compound 1), or a pharmaceutically acceptable salt thereof, in anamount sufficient to provide a reduction in blast cells, e.g., leukemicblast cells, e.g., myeloblasts or myeloid blasts, to thereby treat thedisorder.

In some embodiments, the disorder is an advanced hematologic malignancy,e.g., an advanced hematologic malignancy characterized by the presenceof a mutant allele of IDH1. In some embodiments, the advancedhematologic malignancy is characterized by a mutant allele of IDH1,wherein the IDH1 mutation results in a new ability of the enzyme tocatalyze the NAPH-dependent reduction of α-ketoglutarate toR(−)-2-hydroxyglutarate (2HG) in a patient. In one embodiment, themutant IDH1 has an R132X mutation. In one embodiment, the R132X mutationis selected from R132H, R132C, R132L, R132V, R132S and R132G. In anotheraspect, the R132X mutation is R132H or R132C. In one embodiment, theR132X mutation is R132H.

In some embodiments, the disorder is selected from acute myelogenousleukemia (AML), myelodysplastic syndrome (MDS), myeloproliferativeneoplasms (MPN), myeloproliferative neoplasms (MPN), chronicmyelomonocytic leukemia (CMML), B-acute lymphoblastic leukemias (B-ALL),B-acute lymphoblastic leukemias (B-ALL), and lymphoma (e.g., T-celllymphoma), wherein each is characterized by the presence of a mutantallele of IDH1. In some embodiments, the disorder is selected fromadvanced IDH1 mutation-positive relapsed and/or refractory AML (R/RAML), untreated AML, and MDS.

In some embodiments, the subject has been previously treated with one ormore chemotherapeutic agent(s). In some embodiments, thechemotherapeutic agent is selected from cytarabine (Ara-C),daunorubicin, etoposide, mitoxantrone, idarubicin, 5-azacytidine,decitabine, SGN33A, sargramostim, WT-1 analog peptide vaccine,tipifarnib, MK-8242, campath, and 6 Mercaptopurine (6MP).

In some embodiments, the reduction in blast cells, e.g., leukemic blastcells, e.g., myeloblasts or myeloid blasts, is by about at least afactor of 10, e.g., relative to a reference standard, e.g., by about atleast a factor of 11, e.g., by about at least a factor of 12, e.g., byabout at least a factor of 13, e.g., by about at least a factor of 14,e.g., by about at least a factor of 15, e.g., by about at least a factorof 16, e.g., by about at least a factor of 17, e.g., by about at least afactor of 18, e.g., by about at least a factor of 19, e.g., by about atleast a factor of 20, relative to a reference standard.

In another embodiment, the blast cells, e.g., leukemic blast cells,e.g., myeloblasts or myeloid blasts, are reduced relative to a referencestandard, e.g., to a level that is less than about 10%, e.g., less thanabout 9%, e.g., less than about 8%, e.g., less than about 7%, e.g., lessthan about 6%, e.g., less than about 5%, e.g., less than about 4%, e.g.,less than about 3%, e.g., less than about 2%, e.g., complete remission(CR), relative to a reference standard.

In some embodiments, the reference standard is the level of blast cells,e.g., leukemic blast cells, e.g., myeloblasts or myeloid blasts, in thesubject prior to administration of Compound 1, e.g., in an untreatedsubject, e.g., in a subject not previously treated with Compound 1. Insome embodiments, the subject has been previously treated with one ormore chemotherapeutic agent(s). In some embodiments, thechemotherapeutic agent is selected from cytarabine (Ara-C),daunorubicin, etoposide, mitoxantrone, idarubicin, 5-azacytidine,decitabine, SGN33A, sargramostim, WT-1 analog peptide vaccine,tipifarnib, MK-8242, campath, and 6 Mercaptopurine (6MP).

In some embodiments, the reference standard is the total number of cellsin the sample. In some embodiments, the sample comprises blast cells,myelocytes, neutrophils, promyelocytes, metamyelocytes, and monocytes.

In some embodiments, the subject is monitored for an adverse event. Insome embodiments, the adverse event, includes without limitation,febrile neutropenia, dyspnea, hypotension, mental status changes,neutropenia, increase in the level of blood uric acid, bronchopulmonaryaspergilliosis, dizziness, prolonged electrocardiogram QT, fatigue,intracranial hemorrhage, hypoxia, leukocytosis, leukostasis, lunginfection, metabolic acidosis, nausea, organ failure, pericardialeffusion, fungal pneumonia, pyrexia, renal impairment, retinoic acidsyndrome, septic shock, systemic candida, tachycardia, and vertigo.

In some embodiments, the adverse event is differentiation syndromewherein symptoms comprise fever and/or dyspnea. In some embodiments, thesubject is monitored for differentiation syndrome, and if the subjectexperiences differentiation syndrome is treated with steroids.

In some embodiments, the subject is monitored for an adverse event,e.g., a serious adverse event (SAE), and if an adverse event, e.g., SAE,is experienced by the patient, then treatment is modified ordiscontinued.

Treatment methods described herein can additionally comprise variousevaluation steps prior to and/or following treatment with Compound 1. Insome embodiments, prior to and/or after treatment with Compound 1, themethod further comprises the step of evaluating PK and PD parameters(e.g., tissue, blood, plasma and/or urine concentration(s) of Compound 1or 2HG). This evaluation may be achieved by sample analysis of bodilytissue or bodily fluid, such as blood, plasma or urine by e.g., massspectroscopy, e.g. LC-MS.

Also disclosed herein are solid dispersions, comprising an inhibitor ofmutant IDH1, or a pharmaceutically acceptable salt thereof, and one ormore polymer(s). Also disclosed herein are processes for preparing suchsolid dispersions. These solid dispersions have improved solubility andenhance the exposure of the therapeutically active compound relative toneat crystalline forms of the therapeutically active compound.

Also disclosed herein is the pharmaceutical use of these soliddispersions for treating advanced hematologic malignancies, such asacute myelogenous leukemia (AML), myelodysplastic syndrome (MDS),myeloproliferative neoplasms (MPN), myeloproliferative neoplasms (MPN),chronic myelomonocytic leukemia (CMML), B-acute lymphoblastic leukemias(B-ALL), or lymphoma (e.g., T-cell lymphoma), each characterized by thepresence of a mutant allele of IDH1.

Also disclosed herein are pharmaceutical compositions, comprising thesolid dispersion, and at least one pharmaceutically acceptable carrier.Also disclosed herein are processes for preparing the pharmaceuticalcompositions.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an X-ray powder diffractogram (XRPD) of Form 1.

FIG. 2 is a differential scanning calorimetry (DSC) profile of Form 1.

FIG. 3 is a thermal gravimetric analysis (TGA) profile of Form 1.

FIG. 4 is an X-ray powder diffractogram (XRPD) of Form 2.

FIG. 5 is a differential scanning calorimetry (DSC) profile of Form 2.

FIG. 6 is a thermal gravimetric analysis (TGA) profile of Form 2.

FIG. 7A depicts a line graph showing reduction in 2HG following a singledose (50 mg/kg) of a compound(S)-N-((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide(Compound 1) in an IDH1 mutant R132H xenograft model.

FIG. 7B depicts a bar graph showing Compound 1 (at 0.5 uM, 1 uM and 5 uMconcentrations) reduced intracellular 2HG in primary human IDH-mutatedblast cells (ex vivo).

FIG. 8A depicts a bar graph showing the PK profile following oraladministration of Compound 1 in patients treated at day −3 with a singledose, at day 15 of cycle 1, and at day 1 of cycle 2, each at doses of100 mg BID, 300 mg QD or 500 mg QD.

FIG. 8B depicts a bar graph showing that plasma concentrations of 2HGwere reduced to normal ranges at day −3 with a single dose, at day 15 ofcycle 1, and at day 1 of cycle 2, each at doses of 100 mg BID, 300 mg QDor 500 mg QD.

FIGS. 9A, 9B and 9C depict images of bone marrow aspirate showingblasts, myelocytes, neutrophils, promyelocytes, metamyelocytes andmonocytes, in a patient at baseline, e.g., untreated, after cycle 1, day15 and cycle 1, day 28, respectively.

DETAILED DESCRIPTION OF THE INVENTION

The details of construction and the arrangement of components set forthin the following description or illustrated in the drawings are notmeant to be limiting. Other embodiments and different ways to practicethe invention are expressly included. Also, the phraseology andterminology used herein is for the purpose of description and should notbe regarded as limiting. The use of “including,” “comprising,” or“having,” “containing”, “involving”, and variations thereof herein, ismeant to encompass the items listed thereafter and equivalents thereofas well as additional items.

Definitions

As used above, and throughout the description of the invention, thefollowing terms, unless otherwise indicated, shall be understood to havethe following meanings.

As used herein, the term “acquire” or “acquiring” refers to obtainingpossession of a physical entity (e.g., a sample, e.g., blood sample orblood plasma sample), or a value, e.g., a numerical value, by “directlyacquiring” or “indirectly acquiring” the physical entity or value.“Directly acquiring” means performing a process (e.g., an analyticalmethod) to obtain the physical entity or value. “Indirectly acquiring”refers to receiving the physical entity or value from another party orsource (e.g., a third party laboratory that directly acquired thephysical entity or value). Directly acquiring a value includesperforming a process that includes a physical change in a sample oranother substance, e.g., performing an analytical process which includesa physical change in a substance, e.g., a sample, performing ananalytical method, e.g., a method as described herein, e.g., by sampleanalysis of bodily fluid, such as blood or plasma by, e.g., massspectroscopy, e.g. LC-MS.

As used herein, “crystalline” refers to a solid having a highly regularchemical structure. In particular, a crystalline free base or salt formmay be produced as one or more single crystalline forms. For thepurposes of this application, the terms “crystalline form”, “singlecrystalline form” and “polymorph” are synonymous; the terms distinguishbetween crystals that have different properties (e.g., different XRPDpatterns and/or different DSC scan results). The term “polymorph”includes pseudopolymorphs, which are typically different solvates of amaterial, and thus their properties differ from one another. Thus, eachdistinct polymorph and pseudopolymorph of a free base or salt form isconsidered to be a distinct single crystalline form herein.

The term “substantially crystalline” refers to forms that may be atleast a particular weight percent crystalline. Particular weightpercentages are 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%,99.9%, or any percentage between 10% and 100%. In some embodiments,substantially crystalline refers to a free base or salt form that is atleast 70% crystalline. In other embodiments, substantially crystallinerefers to a free base or salt form that is at least 90% crystalline.

“Form 1” or “compound 1 Form 1” may be used interchangeably, anddescribe the crystalline form synthesized in Example 2, in the Examplessection below, and as described below, and represented by data shown inFIGS. 1, 2, and 3.

“Form 2” or “compound 1 Form 2” are used interchangeably, and describethe crystalline form synthesized in Example 3, in the Examples sectionbelow, and as described below, and represented by data shown in FIGS. 4,5, and 6.

As used herein, “amorphous” refers to a solid material having no longrange order in the position of its atoms. Amorphous solids are generallysupercooled liquids in which the molecules are arranged in a randommanner so that there is no well-defined arrangement and no long rangeorder. Amorphous solids are generally isotropic, i.e., exhibit similarproperties in all directions and do not have definite melting points.For example, an amorphous material is a solid material having no sharpcharacteristic crystalline peak(s) in its X-ray powder diffraction(XRPD) pattern (i.e., is not crystalline as determined by XRPD).Instead, one or several broad peaks (e.g., halos) appear in its XRPDpattern. Broad peaks are characteristic of an amorphous solid. Anamorphous preparation of a compound described herein is substantiallyfree of impurities and/or crystalline compound.

The term “substantially free” refers to forms and compositions that maybe at least a particular weight percent free of impurities and/orcrystalline compound. Particular weight percentages are 60%, 70%, 75%,80%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,99%, 99.5%, 99.9%, or any percentage between 60% and 100% free ofimpurities and/or crystalline compound. In some embodiments,substantially free refers to a free base or salt form that is at least70% pure. In other embodiments, substantially crystalline refers to afree base or salt form that is at least 90% pure. In other embodiments,substantially free of crystalline compound refers to a compositionhaving less than about 30%, less than about 20%, less than about 15%,less than about 10%, less than about 5%, less than about 1% ofcrystalline compound.

As used herein, the terms “isolated” refers to forms that may be atleast a particular weight percent of a particular crystalline form of acompound. Particular weight percentages are 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or any percentage between 90% and100%.

The term “solvate or solvated” means a physical association of acompound, including a crystalline form thereof, of this invention withone or more solvent molecules. This physical association includeshydrogen bonding. In certain instances the solvate will be capable ofisolation, for example when one or more solvent molecules areincorporated in the crystal lattice of the crystalline solid. “Solvateor solvated” encompasses both solution-phase and isolable solvates.Representative solvates include, for example, a hydrate, ethanolates ora methanolate.

The term “hydrate” is a solvate wherein the solvent molecule is H₂O thatis present in a defined stoichiometric amount, and may, for example,include hemihydrate, monohydrate, dihydrate, or trihydrate.

The term “mixture” is used to refer to the combined elements of themixture regardless of the phase-state of the combination (e.g., liquidor liquid/crystalline).

The term “seeding” is used to refer to the addition of a crystallinematerial to initiate recrystallization or crystallization.

The term “antisolvent” is used to refer to a solvent in which compounds,including crystalline forms thereof, are poorly soluble.

As used herein, the term “about” means approximately, in the region of,roughly, or around. When the term “about” is used in conjunction with anumerical range, it modifies that range by extending the boundariesabove and below the numerical values set forth. In general, the term“about” is used herein to modify a numerical value above and below thestated value by a variance of 10%.

As used herein, the term “elevated levels of 2HG” means 10%, 20% 30%,50%, 75%, 100%, 200%, 500% or more 2HG than is present in a subject thatdoes not carry a mutant IDH1 allele. The term “elevated levels of 2HG”may refer to the amount of 2HG within a cell, within a tumor, within anorgan comprising a tumor, or within a bodily fluid.

The term “bodily fluid” includes one or more of amniotic fluidsurrounding a fetus, aqueous humour, blood (e.g., blood plasma), serum,Cerebrospinal fluid, cerumen, chyme, Cowper's fluid, female ejaculate,interstitial fluid, lymph, breast milk, mucus (e.g., nasal drainage orphlegm), pleural fluid, pus, saliva, sebum, semen, serum, sweat, tears,urine, vaginal secretion, or vomit.

As used herein, the terms “inhibit” or “prevent” include both completeand partial inhibition and prevention. An inhibitor may completely orpartially inhibit the intended target.

The term “treat” means decrease, suppress, attenuate, diminish, arrest,or stabilize the development or progression of a disease/disorder (i.e.,an advanced solid tumor, such as acute myelogenous leukemia (AML),myelodysplastic syndrome (MDS), myeloproliferative neoplasms (MPN),myeloproliferative neoplasms (MPN), chronic myelomonocytic leukemia(CMML), B-acute lymphoblastic leukemias (B-ALL), or lymphoma (e.g.,T-cell lymphoma), each characterized by the presence of a mutant alleleof IDH1), lessen the severity of the disease/disorder (i.e., an advancedsolid tumor, such as acute myelogenous leukemia (AML), myelodysplasticsyndrome (MDS), myeloproliferative neoplasms (MPN), myeloproliferativeneoplasms (MPN), chronic myelomonocytic leukemia (CMML), B-acutelymphoblastic leukemias (B-ALL), or lymphoma (e.g., T-cell lymphoma),each characterized by the presence of a mutant allele of IDH1) orimprove the symptoms associated with the disease/disorder (i.e., anadvanced solid tumor, such as acute myelogenous leukemia (AML),myelodysplastic syndrome (MDS), myeloproliferative neoplasms (MPN),myeloproliferative neoplasms (MPN), chronic myelomonocytic leukemia(CMML), B-acute lymphoblastic leukemias (B-ALL), or lymphoma (e.g.,T-cell lymphoma), each characterized by the presence of a mutant alleleof IDH1.

As used herein, an amount of a compound effective to treat a disorder,or a “therapeutically effective amount” refers to an amount of thecompound, which is effective, upon single or multiple doseadministration to a subject, in treating a cell, or in curing,alleviating, relieving or improving a subject with a disorder beyondthat expected in the absence of such treatment.

As used herein, “% w/w” is used to mean by weight as a percentage of atotal weight that is used as the basis for calculating the weightpercentage of an individual component. By way of example, for a bulkcomposition, the % w/w of an individual component may be calculated as apercentage of the total weight of all of the components of the bulkcomposition. By way of another example, for a single oral dosage form,the % w/w of an individual component may be calculated as a percentageof the total weight of all of the components of the single oral dosageform. For example, when the single oral dosage form is a tablet, thetotal weight may be the total weight of all the components of thetablet.

As used herein, the term “subject” is intended to mean human. Exemplaryhuman subjects include a human patient (referred to as a patient) havinga disorder, e.g., a disorder described herein or a normal subject.

The term “physically stable,” as used herein, means that a particularfree base or salt form does not change into one or more differentphysical forms (e.g., different solid forms as measured by XRPD, DSC,etc.) when subjected to specified conditions, e.g., room temperatureambient humidity or 40° C./75% relative humidity, for a specified periodof time, e.g., 1 day, 2 days, 3 days, 1 week, 2 weeks, 1 month, 2months, 3 months, 6 months, 12 months, 18 months, 24 months, or longer.In some embodiments, less than 25% of the form of a compound changesinto one or more different physical forms when subjected to specifiedconditions. In some embodiments, less than about 20%, less than about15%, less than about 10%, less than about 5%, less than about 3%, lessthan about 1%, less than about 0.5% of the form of a particular compoundchanges into one or more different physical forms of that particularcompound when subjected to specified conditions. In some embodiments, nodetectable amount of the particular form of a compound changes into oneor more different physical forms of the compound.

The term “chemically stable,” as used herein, means that the chemicalstructure of a particular compound, does not change into anothercompound (e.g., decompose) when subjected to specified conditions, e.g.,room temperature ambient humidity or 40° C./75% relative humidity, for aspecified period of time, e.g., 1 day, 2 days, 3 days, 1 week, 2 weeks,1 month, 2 months, 3 months, 6 months, 12 months, 18 months, 24 months,or longer. In some embodiments, less than 25% of the form of aparticular compound changes into one or more other compounds whensubjected to specified conditions. In some embodiments, less than about20%, less than about 15%, less than about 10%, less than about 5%, lessthan about 3%, less than about 1%, less than about 0.5% of the form of aparticular compound changes into one or more other compounds whensubjected to specified conditions. In some embodiments, no detectableamount of the form of a particular compound changes into one or moredifferent physical forms of that particular compound.

The term “dispersion” refers to a disperse system in which onesubstance, the dispersed phase, is distributed, in discrete units,throughout a second substance (the continuous phase or vehicle). Thesize of the dispersed phase can vary considerably (e.g., colloidalparticles of nanometer dimension, to multiple microns in size). Ingeneral, the dispersed phases can be solids, liquids, or gases. In thecase of a solid dispersion, the dispersed and continuous phases are bothsolids. In pharmaceutical applications, a solid dispersion can include acrystalline therapeutically active compound (dispersed phase) in anamorphous polymer(s) (continuous phase), or alternatively, an amorphoustherapeutically active compound (dispersed phase) in an amorphouspolymer (continuous phase).

The term “amorphous solid dispersion” generally refers to a soliddispersion of two or more components, usually a therapeutically activecompound and polymer (or plurality of polymers), but possibly containingother components such as surfactants or other pharmaceutical excipients,where the therapeutically active compound is in the amorphous phase, andthe physical stability and/or dissolution and/or solubility of theamorphous therapeutically active compound is enhanced by the othercomponents. In some embodiments, an amorphous solid dispersion includesthe polymer(s) (and optionally a surfactant) constituting the dispersedphase, and the therapeutically active compound constitutes thecontinuous phase. In some embodiments, an amorphous solid dispersionincludes the polymer(s) (and optionally a surfactant) constituting thecontinuous phase, and the therapeutically active compound constitutesthe dispersed phase.

An exemplary solid dispersion is a co-precipitate or a co-melt of aparticular therapeutically active compound with one or more polymer(s).A “co-precipitate” is produced after dissolving a therapeutically activecompound and one or more polymer(s) in a solvent or solvent mixturefollowed by the removal of the solvent or solvent mixture. Sometimes theone or more polymer(s) can be suspended in the solvent or solventmixture. The solvent or solvent mixture includes organic solvents andsupercritical fluids. The solvent or solvent mixture can also contain anon-volatile solvent. A “co-melt” is produced after heating atherapeutically active compound and one or more polymer(s) to melt,optionally in the presence of a solvent or solvent mixture, followed bymixing, removal of at least a portion of the solvent if applicable, andcooling to room temperature at a selected rate. In some cases, soliddispersions are prepared by adding a solution of a therapeuticallyactive compound and solid polymers followed by mixing and removal of thesolvent or solvent mixture. To remove the solvent or solvent mixture,vacuum drying, spray drying, tray drying, lyophilization, and otherdrying procedures may be applied. Applying any of these methods usingappropriate processing parameters, according to this disclosure, wouldprovide the particular therapeutically active compound in an amorphousstate in the final solid dispersion product.

As used herein, the term “directly compressed dosage form” generallyrefers to a form (e.g., a tablet) that is obtained by the compression ofa dry blend of powders (e.g., solid dispersion, e.g., agglomerateddispersion) that comprise a compound, e.g., a therapeutic compound(e.g., a poorly soluble therapeutic compound, e.g., compound 1, e.g.,amorphous compound 1, e.g., in a solid dispersion, e.g., that alsoincludes one or more polymer(s) and optionally one or moresurfactant(s)) and optionally one or more excipients. For example, theproduct (e.g., solid dispersion) resulting from a process describedherein can have improved properties (e.g., flowability) that allow it tobe directly compressed, e.g., into an oral dosage form, e.g., tablets,or to be formulated into capsules or saches.

Pharmaceutical Compositions and Methods of Treatment

Provided is a method of treating advanced solid tumors, such as acutemyelogenous leukemia (AML), myelodysplastic syndrome (MDS),myeloproliferative neoplasms (MPN), myeloproliferative neoplasms (MPN),chronic myelomonocytic leukemia (CMML), B-acute lymphoblastic leukemias(B-ALL), or lymphoma (e.g., T-cell lymphoma), each characterized by thepresence of a mutant allele of IDH1 comprising administering to asubject in need thereof a pharmaceutical composition comprising: (a) acompound(S)-N-((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide(Compound 1), or a pharmaceutically acceptable salt thereof, as part ofa solid dispersion, and optionally (b) one or more pharmaceuticallyacceptable carrier(s).

Also provided are compositions containing Compound 1, or apharmaceutically acceptable salt thereof, as part of a solid dispersion(e.g., an amorphous solid dispersion). Also provided are pharmaceuticalcompositions, comprising: (a) Compound 1, or a pharmaceuticallyacceptable salt thereof, as part of a solid dispersion, and (b) one ormore pharmaceutically acceptable carrier(s).

These methods of treatment and pharmaceutical compositions are furtherillustrated by the detailed descriptions and illustrative examples givenbelow.

Pharmaceutical compositions comprising solid dispersions of atherapeutically active compound in a matrix can provide improvedchemical and physical properties and can be prepared by forming ahomogeneous solution or melt of the therapeutically active compound andmatrix material followed by solidifying the mixture by cooling, orremoval of the solvent. Such solid dispersions of therapeutically activecompounds often show enhanced bioavailability when administered orallyrelative to oral compositions comprising the undispersed compound.

Spray drying is the most widely used industrial process involvingparticle formation and drying, and can be used to produce soliddispersions of therapeutically active compounds. It is highly suited forthe continuous production of dry solids in either powder, granulate oragglomerate form from liquid feedstocks as solutions, emulsions andpumpable suspensions. Therefore, spray drying is a useful process wherethe end-product must comply with precise quality standards regardingparticle size distribution, residual moisture content, bulk density, andparticle shape.

Critical quality attributes of a spray-dried dispersion include potency,related substances, residual solvent content, homogeneity, lack ofcrystallinity, dissolution performance, particle morphology, and bulkpowder flow properties.

Critical process parameters include spray solution composition andviscosity, nozzle type and dimensions, atomization pressure, spraysolution feed rate, drying gas flow rate, inlet and outlet temperatures,condenser temperature (e.g., for closed-loop drying processes), andsecondary drying parameters.

In one embodiment, at least a particular percentage by weight ofCompound 1 is crystalline. Particular weight percentages may be 10%,20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or any percentagebetween 10% and 100%. When a particular percentage by weight of Compound1 is crystalline, the remainder of Compound 1 is the amorphous form ofCompound 1. Non-limiting examples of crystalline Compound 1 include asingle crystalline form of Compound 1 or a mixture of different singlecrystalline forms. In some embodiments, Compound 1 is at least 90% byweight crystalline. In some other embodiments, Compound 1 is at least95% by weight crystalline. In some other embodiments, Compound 1 is atleast 99% by weight crystalline.

In another embodiment, a particular percentage by weight of thecrystalline Compound 1 is a specific single crystalline form or acombination of single crystalline forms. Particular weight percentagesmay be 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or anypercentage between 10% and 100%. In another embodiment, Compound 1 is atleast 90% by weight of a single crystalline form. In another embodiment,Compound 1 is at least 95% by weight of a single crystalline form. Inanother embodiment, Compound 1 is at least 99% by weight of a singlecrystalline form.

In the following description of Compound 1, embodiments of the inventionmay be described with reference to a particular crystalline form ofCompound 1, as characterized by one or more properties as discussedherein. The descriptions characterizing the crystalline forms may alsobe used to describe the mixture of different crystalline forms that maybe present in a crystalline Compound 1. However, the particularcrystalline forms of Compound 1 may also be characterized by one or moreof the characteristics of the crystalline form as disclosed herein, withor without regard to referencing a particular crystalline form.

The crystalline forms are further illustrated by the detaileddescriptions and illustrative examples given below. The XRPD peaksdescribed in Tables 1 and 2 may vary by 0.2 depending upon theinstrument used to obtain the data.

Form 1

In one embodiment, a single crystalline form, Form 1, of the compound 1is characterized by the X-ray powder diffraction (XRPD) pattern shown inFIG. 1, and data shown in Table 1, obtained using CuKa radiation. In aparticular embodiment, the polymorph can be characterized by one or moreof the peaks taken from FIG. 1, as shown in Table 1. For example, thepolymorph can be characterized by one or two or three or four or five orsix or seven or eight or nine of the peaks shown in Table 1.

TABLE 1 Angle 2-Theta ° Intensity % 8.6 90.3 13.2 60.0 15.6 85.5 18.572.5 19.6 31.5 20.6 71.6 21.6 100.0 26.4 64.2 27.3 45.6

In another embodiment, Form 1 can be characterized by the peaksidentified at 2θ angles of 8.6, 15.6, 18.5, 20.6, 21.6, and 26.4°. Inanother embodiment, Form 1 can be characterized by the peaks identifiedat 2θ angles of 8.6, 15.6, 18.5, and 21.6°.

In another embodiment, Form 1 can be characterized by the differentialscanning calorimetry profile (DSC) shown in FIG. 2. The DSC graph plotsthe heat flow as a function of temperature from a sample, thetemperature rate change being about 10° C./min. The profile ischaracterized by an endothermic transition with an onset temperature ofabout 140.1° C. with a melt at about 149.9° C.

In another embodiment, Form 1 can be characterized by thermalgravimetric analysis (TGA) shown in FIG. 3. The TGA profile graphs thepercent loss of weight of the sample as a function of temperature, thetemperature rate change being about 10° C./min. The weight lossrepresents a loss of about 0.44% of the weight of the sample as thetemperature is changed from about 29.0° C. to 125.0° C.

Form 2

In one embodiment, a single crystalline form, Form 2, of the compound 1is characterized by the X-ray powder diffraction (XRPD) pattern shown inFIG. 4, and data shown in Table 2, obtained using CuKa radiation. In aparticular embodiment, the polymorph can be characterized by one or moreof the peaks taken from FIG. 4, as shown in Table 2. For example, thepolymorph can be characterized by one or two or three or four or five orsix or seven or eight or nine or ten of the peaks shown in Table 2.

TABLE 2 Angle 2-Theta ° Intensity % 9.8 85.6 11.6 100.0 14.9 11.4 16.515.3 19.6 75.2 20.1 7.3 22.5 32.6 23.0 69.4 25.0 8.9 31.4 22.0

In another embodiment, Form 2 can be characterized by the peaksidentified at 2θ angles of 9.8, 11.6, 19.6, 22.5, 23.0, and 31.4°. Inanother embodiment, Form 2 can be characterized by the peaks identifiedat 2θ angles of 9.8, 11.6, 19.6, and 23.0°.

In another embodiment, Form 2 can be characterized by the differentialscanning calorimetry profile (DSC) shown in FIG. 5. The DSC graph plotsthe heat flow as a function of temperature from a sample, thetemperature rate change being about 10° C./min. The profile ischaracterized by an endothermic transition with an onset temperature ofabout 62.7° C. with a melt at about 72.5° C., and an endothermictransition with an onset temperature of about 145.6° C. with a melt atabout 153.6° C.

In another embodiment, Form 2 can be characterized by thermalgravimetric analysis (TGA) shown in FIG. 6. The TGA profile graphs thepercent loss of weight of the sample as a function of temperature, thetemperature rate change being about 10° C./min. The weight lossrepresents a loss of about 0.57% of the weight of the sample as thetemperature is changed from about 29.3° C. to 170.3° C.

Other embodiments are directed to a single crystalline form of compound1 characterized by a combination of the aforementioned characteristicsof any of the single crystalline forms discussed herein. Thecharacterization may be by any combination of one or more of the XRPD,TGA, and DSC described for a particular polymorph. For example, thesingle crystalline form of compound 1 may be characterized by anycombination of the XRPD results regarding the position of the majorpeaks in a XRPD scan; and/or any combination of one or more ofparameters derived from data obtained from a XRPD scan. The singlecrystalline form of compound 1 may also be characterized by TGAdeterminations of the weight loss associated with a sample over adesignated temperature range; and/or the temperature at which aparticular weight loss transition begins. DSC determinations of thetemperature associated with the maximum heat flow during a heat flowtransition and/or the temperature at which a sample begins to undergo aheat flow transition may also characterize the crystalline form. Weightchange in a sample and/or change in sorption/desorption of water permolecule of Compound 1 as determined by water sorption/desorptionmeasurements over a range of relative humidity (e.g., 0% to 90%) mayalso characterize a single crystalline form of Compound 1.

Solid Dispersions

Provided are compositions, comprising Compound 1, or a pharmaceuticallyacceptable salt thereof, and one or more polymer(s) as part of a soliddispersion (e.g., an amorphous solid dispersion). In some embodiments,the solid dispersion comprises Compound 1, or a pharmaceuticallyacceptable salt thereof, and one or more polymer(s). In someembodiments, the solid dispersion comprises Compound 1, or apharmaceutically acceptable salt thereof, one or more polymer(s), andone or more surfactant(s). In some embodiments, the solid dispersioncomprises Compound 1, or a pharmaceutically acceptable salt thereof, andone polymer. In some embodiments, the solid dispersion comprisesCompound 1, or a pharmaceutically acceptable salt thereof, one polymer,and a surfactant.

The solid dispersions provided herein, comprising Compound 1, or apharmaceutically acceptable salt thereof, can enhance the solubility ofCompound 1 relative to a neat crystalline form of Compound 1 (e.g., Form1 or Form 2), and thus provide improved exposure upon oral dosing of thesolid dispersion to a subject. In one embodiment, the solid dispersioncomprises Compound 1, or a pharmaceutically acceptable salt thereof, oneor more polymer(s), and optionally one or more solubility enhancingsurfactant.

For example, the aqueous solubility of Form 1 is about 0.025 mg/mL toabout 0.035 mg/mL and the aqueous solubility of Form 2 is about 0.008mg/mL to about 0.010 mg/mL.

Form 2 has a solubility of about 0.018 mg/mL in fasted state simulatedintestinal fluid (FASSIF) at a pH of 6.1 at 4 hours. In comparison,amorphous spray-dried dispersions have a solubility of about 0.05 mg/mLto about 0.50 mg/mL in FASSIF at 3 hours.

In some embodiments, the solid dispersion exhibits at least about 20%,at least about 30%, at least about 40%, at least about 50%, at leastabout 60%, at least about 70%, at least about 80%, or at least about 90%higher exposure of Compound 1, or a pharmaceutically acceptable saltthereof, when administered to a subject as compared to administration ofin-situ amorphous Compound 1, or a pharmaceutically acceptable saltthereof. In some embodiments, the solid dispersion exhibits at leastabout 20%, at least about 30%, at least about 40%, at least about 50%,at least about 60%, at least about 70%, at least about 80%, or at leastabout 90% higher exposure of Compound 1, or a pharmaceuticallyacceptable salt thereof, when administered to a subject as compared toadministration of neat crystalline Compound 1, or a pharmaceuticallyacceptable salt thereof.

In rat and monkey pharmacokinetics studies, modest exposure improvementis observed upon administration of solid dispersion oral dosage forms ascompared to in-situ amorphous dosing shows. For example, a soliddispersion containing 50% w/w Compound 1 and 50% w/w Polyvinyl AcetatePhthalate (PVAP) has approximately two-fold higher exposure as comparedto in-situ amorphous Compound 1 in male Sprague Dawley rats. There is nosignificant difference in exposure between a solid dispersion containing70% w/w Compound 1 and 30% w/w oral dosage form as compared to in-situamorphous Compound 1. In male cynomolgus monkeys, the exposure of asolid dispersion containing 50% w/w Compound 1 and 50% w/whydroxypropylmethylcellulose acetate succinate, also known ashpromellose acetate succinate, (HPMCAS) shows no significant differenceas compared to the in-situ amorphous Compound 1. Similarly, a soliddispersion containing 50% w/w Compound 1 and 50% w/whydroxypropylmethylcellulose also known as hypromellose phthalate(HPMC-Phthalate) shows no significant difference as compared to thein-situ amorphous Compound 1. While in-situ amorphous therapeuticcompounds are commonly used for dosing in animal studies, they are notsuitable dosage forms for dosing in humans.

As described in the rat pharmacokinetics study of Example 4, Compound 1exposure is improved when solid dispersion dosage forms are administeredas compared to neat crystalline Compound 1 Form 2.

In some embodiments, at least a portion of Compound 1, or apharmaceutically acceptable salt thereof, in the solid dispersion is inthe amorphous state (e.g., at least about 50%, at least about 55%, atleast about 60%, at least about 65%, at least about 70%, at least about75%, at least about 80%, at least about 85%, at least about 90%, atleast about 95%, at least about 98%, or at least about 99%). In otherembodiments, the solid dispersion is substantially free of crystallineCompound 1, or a pharmaceutically acceptable salt thereof.

In some embodiments, the composition is an amorphous solid (e.g. spraydried) dispersion comprising Compound 1, or a pharmaceuticallyacceptable salt thereof, and a polymer. The amorphous solid dispersioncan include, e.g., less than about 30%, less than about 20%, less thanabout 15%, less than about 10%, less than about 5%, less than about 4%,less than about 3%, less than about 2%, or less than about 1% ofcrystalline Compound 1, or a pharmaceutically acceptable salt thereof,e.g., be substantially free of crystalline Compound 1, or apharmaceutically acceptable salt thereof.

In one embodiment, the solid dispersion exhibits a predetermined levelof physical and/or chemical stability. E.g., the solid dispersionretains about 50%, about 60%, about 70%, about 80%, about 90%, about95%, about 98%, or about 99%, of amorphous Compound 1, or apharmaceutically acceptable salt thereof, when stored at 25° C. in aclosed water tight container, e.g., an amber glass vial, high densitypolyethylene (HDPE) container or double polyethylene bags with twistednylon tie placed in an HDPE container with desiccant.

In some embodiments, the polymer increases the chemical or physicalstability (e.g., as measured by a Modulated Differential ScanningCalorimeter) of Compound 1, or a pharmaceutically acceptable saltthereof, when stored (e.g., at 2-8° C., e.g. 4° C. or at roomtemperature) by at least about 10% (e.g., by at least about 20%, by atleast about 30%, by at least about 40%, by at least about 50%, by atleast about 60%, by at least about 70%, by at least about 80%, or by atleast about 90%) compared to amorphous Compound 1, or a pharmaceuticallyacceptable salt thereof, without being in the presence of the polymer.

A solid dispersion generally exhibits a glass transition temperature,where the dispersion makes a transition from a glassy solid to a rubberycomposition. In general, the higher the glass transition temperature,the greater the physical stability of the dispersion. The existence of aglass transition temperature generally indicates that at least a largeportion of the composition (e.g., dispersion) is in an amorphous state.The glass transition temperature (Tg) of a solid dispersion suitable forpharmaceutical applications is generally at least about 50° C. In someembodiments, higher temperatures are preferred. Therefore, in someembodiments, a solid dispersion disclosed herein has a Tg of at leastabout 100° C. (e.g., at least about 100° C., at least about 105° C., atleast about 110° C., at least about 115° C., at least about 120° C., atleast about 125° C., at least about 130° C., at least about 135° C., atleast about 140° C., at least about 150° C., at least about 160° C., atleast about 170° C., at least about 175° C., at least about 180° C., orat least about 190° C.). In some embodiments, the Tg is up to about 200°C. In some embodiments, the Tg is up to about 130° C. (e.g., at leastabout 110° C., at least about 111° C., at least about 112° C., at leastabout 113° C., at least about 114° C., at least about 115° C., at leastabout 116° C., at least about 117° C., at least about 118° C., at leastabout 119° C., at least about 120° C., at least about 121° C., at leastabout 122° C., at least about 123° C., at least about 124° C., at leastabout 125° C., at least about 1216° C., at least about 127° C., at leastabout 128° C., at least about 129° C., or at least about 130° C.).Unless otherwise noted, the glass transition temperatures disclosedherein are measured under dry conditions.

In some embodiments the solid dispersion has a higher glass transitiontemperature than the glass transition temperature of amorphous Compound1, or a pharmaceutically acceptable salt thereof, without being in thepresence of the polymer(s). In some embodiments, the solid dispersionhas a relaxation rate that is lower than the relaxation rate ofamorphous Compound 1, or a pharmaceutically acceptable salt thereof,without being in the presence of the polymer(s).

Examples of polymers in the solid dispersion include cellulosederivatives (e.g., hydroxypropylmethylcellulose also known ashypromellose, (HPMC), hydroxypropylmethylcellulose phthalate, also knownas hypromellose phthalate (HPMCP), hydroxypropylmethylcellulose acetatesuccinate, also known as hpromellose acetate succinate, (HPMCAS),hydroxypropylcellulose (HPC)), ethylcellulose, or cellulose acetatephthalate; polyvinylpyrrolidones (PVP); polyethylene glycols (PEG);polyvinyl alcohols (PVA); polyvinyl esters, such as Polyvinyl AcetatePhthalate (PVAP); acrylates, such as polymethacrylate (e.g., Eudragit®E); cyclodextrins (e.g., .beta.-cyclodextrin); Poly (D, L-lactide)(PLA), Poly (D,L-lactide, co-glycolide acid (PLGA); and copolymers andderivatives thereof, including for example polyvinylpyrollidone-vinylacetate (PVP-VA), Polyvinyl caprolactam-polyvinyl, andacetate-polyethyleneglycol copolymer, Methylacrylate/methacrylic acidcopolymer; Soluplus; Copovidone; and mixtures thereof.

In some embodiments, the solid dispersion includes one water-solublepolymer. In some embodiments, the solid dispersion includes onepartially water-soluble polymer. In some embodiments, the polymer is acellulose polymer.

In some embodiments, the polymer is HPMCAS (e.g., HPMCAS of differentgrades: HPMCAS-M, HPMCAS-MG or HPMCAS-HG). In some embodiments, thepolymer is PVAP. In some embodiments, the polymer is HPMC (e.g., HPMC ofdifferent grades: HMPC60SH50, HPMCE50 or HPMCE15). In some embodiments,the polymer is HPMCP (e.g., HPMCP of different grades: e.g.,HMPCP-HP55).

In some embodiments, the polymer is a pH-dependent enteric polymer. SuchpH-dependent enteric polymers include, but are not limited to, cellulosederivatives (e.g., cellulose acetate phthalate (CAP)), HPMCP, HPMCAS,carboxymethylcellulose (CMC) or a salt thereof (e.g., a sodium salt suchas (CMC-Na)); cellulose acetate trimellitate (CAT),hydroxypropylcellulose acetate phthalate (HPCAP),hydroxypropylmethyl-cellulose acetate phthalate (HPMCAP), andmethylcellulose acetate phthalate (MCAP), polymethacrylates (e.g.,Eudragit S), or mixtures thereof.

In some embodiments, the polymer is hydroxypropylmethylcellulose acetatesuccinate, also known as hypromellose acetate succinate, (HPMCAS), e.g.,HMPCAS-HG.

In another embodiment, the polymer(s) is an insoluble cross-linkedpolymer, for example a polyvinylpyrrolidone (e.g., Crospovidone). Inanother embodiment, the polymer(s) is polyvinylpyrrolidone (PVP).

In some embodiments, the one or more polymer(s) is present in the soliddispersion in an amount of between about 10% w/w and 90% w/w (e.g.,between about 20% w/w and about 80% w/w; between about 30% w/w and about70% w/w; between about 40% w/w and about 60% w/w; or between about 15%w/w and about 35% w/w). In some embodiments, the polymer(s) is presentin the solid dispersion in an amount of from about 10% w/w to about 80%w/w, for example from about 30% w/w to about 75% w/w, or from about 40%w/w to about 65% w/w, or from about 45% w/w to about 55% w/w, forexample, about 46% w/w, about 47% w/w, about 48% w/w, about 49% w/w,about 50% w/w, about 51% w/w, about 52% w/w, about 53% w/w, or about 54%w/w. In some embodiments, the polymer(s) is present in the soliddispersion in an amount of about 48% w/w, about 48.5% w/w, about 49%w/w, about 49.5% w/w, about 50% w/w, about 50.5% w/w, about 51% w/w,about 51.5% w/w, about 52% w/w, or about 52.5% w/w.

In some embodiments, the polymer(s) is present in the solid dispersionin an amount of from about 30% w/w to about 70% w/w. In someembodiments, the polymer(s) is present in the solid dispersion in anamount of from about 35% w/w to about 65% w/w. In some embodiments, thepolymer(s) is present in the solid dispersion in an amount of from about40% w/w to about 60% w/w. In some embodiments, the polymer(s) is presentin the solid dispersion in an amount of from about 45% w/w to about 55%w/w. In some embodiments, the polymer(s) is present in the soliddispersion in an amount of about 50% w/w.

In some embodiments, Compound 1, or a pharmaceutically acceptable saltthereof, is present in the solid dispersion in an amount of from about10% w/w and 90% w/w (e.g., between about 20% w/w and about 80% w/w;between about 30% w/w and about 70% w/w; between about 40% w/w and about60% w/w; or between about 15% w/w and about 35% w/w). In someembodiments, Compound 1, or a pharmaceutically acceptable salt thereof,is present in the solid dispersion in an amount of from about 10% w/w toabout 80% w/w, for example from about 30% w/w to about 75% w/w, or fromabout 40% w/w to about 65% w/w, or from about 45% w/w to about 55% w/w,for example, about 46% w/w, about 47% w/w, about 48% w/w, about 49% w/w,about 50% w/w, about 51% w/w, about 52% w/w, about 53% w/w, or about 54%w/w. In some embodiments, Compound 1, or a pharmaceutically acceptablesalt thereof, is present in the solid dispersion in an amount of about48% w/w, about 48.5% w/w, about 49% w/w, about 49.5% w/w, about 50% w/w,about 50.5% w/w, about 51% w/w, about 51.5% w/w, about 52% w/w, or about52.5% w/w.

In some embodiments, Compound 1, or a pharmaceutically acceptable saltthereof, is present in the solid dispersion in an amount of from about30% w/w to about 70% w/w. In some embodiments, Compound 1, or apharmaceutically acceptable salt thereof, is present in the soliddispersion in an amount of from about 35% w/w to about 65% w/w. In someembodiments, Compound 1, or a pharmaceutically acceptable salt thereof,is present in the solid dispersion in an amount of from about 40% w/w toabout 60% w/w. In some embodiments, Compound 1, or a pharmaceuticallyacceptable salt thereof, is present in the solid dispersion in an amountof from about 45% w/w to about 55% w/w. In some embodiments, Compound 1,or a pharmaceutically acceptable salt thereof, is present in the soliddispersion in an amount of about 50% w/w.

In another embodiment, the solid dispersion includes about 20% w/w toabout 80% w/w Compound 1, or a pharmaceutically acceptable salt thereof,and about 20% w/w to about 80% of polymer(s). In another embodiment, thesolid dispersion includes about 25% w/w to about 75% w/w Compound 1, ora pharmaceutically acceptable salt thereof, and about 25% w/w to about75% of polymer(s). In another embodiment, the solid dispersion includesabout 30% w/w to about 70% w/w Compound 1, or a pharmaceuticallyacceptable salt thereof, and about 30% w/w to about 70% of polymer(s).In another embodiment, the solid dispersion includes about 35% w/w toabout 65% w/w Compound 1, or a pharmaceutically acceptable salt thereof,and about 35% w/w to about 65% of polymer(s). In another embodiment, thesolid dispersion includes about 40% w/w to about 60% w/w Compound 1, ora pharmaceutically acceptable salt thereof, and about 40% w/w to about60% of polymer(s). In another embodiment, the solid dispersion includesabout 45% w/w to about 55% w/w Compound 1, or a pharmaceuticallyacceptable salt thereof, and about 45% w/w to about 55% of polymer(s).In another embodiment, the solid dispersion includes about 50% w/wCompound 1, or a pharmaceutically acceptable salt thereof, and about 50%w/w of polymer(s).

In another embodiment, the solid dispersion includes about 45% w/w toabout 55% w/w Compound 1, or a pharmaceutically acceptable salt thereof,and about 45% w/w to about 55% w/w HPMCAS (e.g., HPMCAS-MG or HPMCAS-HG,or other grades such as LF, MF, HF, or LG) or PVAP. In anotherembodiment, the solid dispersion includes about 50% w/w Compound 1, or apharmaceutically acceptable salt thereof, and about 50% w/w of HPMCAS.

In some embodiments, the solid dispersion also includes a surfactant orinert pharmaceutically acceptable substance. Examples of surfactants inthe solid dispersion include sodium lauryl sulfate (SLS), vitamin E or aderivative thereof (e.g., vitamin E TPGS), Docusate Sodium, sodiumdodecyl sulfate, polysorbates (such as Tween 20 and Tween 80),poloxamers (such as Poloxamer 335 and Poloxamer 407), glycerylmonooleate, Span 65, Span 25, Capryol 90, pluronic copolymers (e.g.,Pluronic F108, Pluronic P-123), and mixtures thereof. In someembodiments, the surfactant is SLS. In some embodiments, the surfactantis vitamin E or a derivative thereof (e.g., vitamin E TPGS).

In some embodiments, the surfactant is present in the solid dispersionin an amount of from about 0.1% w/w to about 10% w/w, for example fromabout 0.5% w/w to about 2% w/w, or from about 1% w/w to about 3% w/w,from about 1% w/w to about 4% w/w, or from about 1% w/w to about 5% w/w.In some embodiments, the surfactant is present in the solid dispersionin an amount of about 0.1% w/w, about 0.2% w/w, about 0.3% w/w, about0.4% w/w, about 0.5% w/w, about 0.6% w/w, about 0.7% w/w, about 0.8%w/w, about 0.9% w/w, or about 1% w/w. In some embodiments, thesurfactant is present in the solid dispersion in an amount of about 0.5%w/w, about 1% w/w, about 1.5% w/w, about 2% w/w, about 2.5% w/w, about3% w/w, about 3.5% w/w, about 4% w/w, about 4.5% w/w, or about 5% w/w.

Processes for Preparing Solid Dispersions

In some embodiments, the solid dispersion may be prepared according to aprocess described herein. In general, methods that could be used includethose that involve rapid removal of solvent or solvent mixture from amixture or cooling a molten sample. Such methods include, but are notlimited to, rotational evaporation, freeze-drying (i.e.,lyophilization), vacuum drying, melt congealing, and melt extrusion. Oneembodiment of this disclosure involves solid dispersion obtained byspray-drying. In one embodiment, the product obtained by spray drying isdried to remove the solvent or solvent mixture.

Preparations disclosed herein, e.g., a pharmaceutical composition, canbe obtained by spray-drying a mixture comprising Compound 1, or apharmaceutically acceptable salt thereof, one or more polymer(s), and anappropriate solvent or solvent mixture. Spray drying involvesatomization of a liquid mixture containing, e.g., a solid and a solventor solvent mixture, and removal of the solvent or solvent mixture. Thesolvent or solvent mixture can also contain a nonvolatile solvent, suchas glacial acetic acid. Atomization may be done, for example, through atwo-fluid or pressure or electrosonic nozzle or on a rotating disk.

Spray drying converts a liquid feed to a dried particulate form. Spraydrying generally involves the atomization of a liquid feed solution intoa spray of droplets and contacting the droplets with hot air or gas in adrying chamber. The sprays are generally produced by either rotary(wheel) or nozzle atomizers. Evaporation of moisture from the dropletsand formation of dry particles proceed under controlled temperature andairflow conditions.

Optionally, a secondary drying process such as fluidized bed drying orvacuum drying, may be used to reduce residual solvents (and otheradditives, such as glacial acetic acid) to pharmaceutically acceptablelevels. Typically, spray-drying involves contacting a highly dispersedliquid suspension or solution (e.g., atomized solution), and asufficient volume of hot air or gas (e.g., nitrogen, e.g., purenitrogen) to produce evaporation and drying of the liquid droplets. Thepreparation to be spray dried can be any solution, coarse suspension,slurry, colloidal dispersion, or paste that may be atomized using theselected spray-drying apparatus. In a standard procedure, thepreparation is sprayed into a current of warm filtered air (or into gas,e.g., nitrogen) that evaporates the solvent and conveys the driedproduct to a collector (e.g., a cyclone). The spent air or gas is thenexhausted with the solvent (or solvent mixture including any additivessuch as glacial acetic acid), (e.g., then filtered) or alternatively thespent air or gas is sent to a condenser to capture and potentiallyrecycle the solvent or solvent mixture. For example, if a gas (e.g.,nitrogen) is used, the gas is then optionally recycled, heated again andreturned to the unit in a closed loop system. Commercially availabletypes of apparatus may be used to conduct the spray-drying. For example,commercial spray dryers are manufactured by Buchi Ltd. and Niro (e.g.,the PSD line of spray driers manufactured by Niro).

Spray-drying typically employs solids loads of material from about 1% toabout 30% or up to about 50% (i.e., therapeutically active Compound plusand excipients), preferably at least about 10%. In some embodiments,solids loads of less than 10% may result in poor yields and unacceptablylong run-times. In general, the upper limit of solids loads is governedby the viscosity of (e.g., the ability to pump) the resulting solutionand the solubility of the components in the solution. Generally, theviscosity of the solution can determine the size of the particle in theresulting powder product.

Techniques and methods for spray-drying may be found in Perry's ChemicalEngineering Handbook, 6th Ed., R. H. Perry, D. W. Green & J. O. Maloney,eds., McGraw-Hill Book Co. (1984); and Marshall “Atomization andSpray-Drying” 50, Chem. Eng. Prog. Monogr. Series 2 (1954). In general,the spray-drying is conducted with an inlet temperature of from about40° C. to about 200° C., for example, from about 70° C. to about 150°C., preferably from about 40° C. to about 60° C., about 50° C. to about55° C., or about 80° C. to about 110° C., e.g., about 90° C. Thespray-drying is generally conducted with an outlet temperature of fromabout 20° C. to about 100° C., for example from about 25° C. to about30° C. (e.g., about 26° C.), about 40° C. to about 50° C., about 50° C.to about 65° C., e.g., about 56° C. to about 58° C.

Removal of the solvent or solvent mixture may require a subsequentdrying step, such as tray drying, fluid bed drying (e.g., from aboutroom temperature to about 100° C.), vacuum drying, microwave drying,rotary drum drying or biconical vacuum drying (e.g., from about roomtemperature to about 200° C.).

In one embodiment, the spray-drying is fluidized spray drying (FSD). Thesteps in FSD can include, for example: preparing a liquid feed solution(e.g., containing Compound 1 or a pharmaceutically acceptable saltthereof, and optionally a polymer(s) and/or surfactant(s), dissolved orsuspended in solvent(s)); atomizing (e.g., with a pressure nozzle, arotary atomizer or disk, two-fluid nozzle or other atomizing methods)the feed solution upon delivery into the drying chamber of a spraydryer, e.g., operating in FSD mode; drying the feed solution in thedrying chamber with heated air or a heated gas (e.g., nitrogen) toobtain a product, wherein larger particles of product separate out,e.g., drop out, while fines are carried by a stream of air or gas up tothe top of the drying chamber (e.g., by natural convection) and to acyclone, and re-introducing (e.g., at the top of the drying chamber oraxially to the middle of the chamber) the fines into the drying chamber,wherein the re-introduced fines can agglomerate with newly formedproduct to generate an agglomerated product, wherein if the agglomeratedproduct is large enough, it will separate out, if it is not large enoughto separate out, the agglomerated product will be carried by convectionto the top of the chamber and to the cyclone and re-introduced into thechamber. This process repeats until an agglomerated product that islarge enough to drop out is formed. The fines can be re-introduced fromthe cyclone to the drying chamber via a feed pipe.

In some embodiments, rather than drying the feed solution with heatedair or a heated gas, the feed solution can instead be spray congealed,e.g., the chamber is at room temperature (e.g., 21±4° C.) or is cooled,e.g., cooled gas (e.g., nitrogen) is used for the process.

FSD can further include collecting the agglomerated product in a firstfluidizing chamber; which can be followed by discharging theagglomerated product from the first fluidizing chamber to a secondfluidizing chamber, wherein a post-drying process can occur.

The agglomerated product (e.g., that separates out in the dryingchamber) can then be transferred from the second fluidizing chamber to athird fluidizing chamber, where the agglomerated product is cooled. Theagglomerated product (e.g., a solid dispersion of an amorphous compound)can then be further processed. For example, the product can be directlycompressed. The product can optionally be blended with a surfactant,excipient, or pharmaceutically acceptable carrier, e.g., prior to directcompression. The product can optionally be further processed, e.g.,milled, granulated, blended, and/or mixed with a melt granulate,surfactant, excipient, and/or pharmaceutically acceptable carrier.

FSD can be performed in a commercial spray dryer operating in fluidizedspray dryer mode (FSD mode). FSD can be accomplished in either opencycle mode or closed cycle mode (e.g., the drying gas, e.g., nitrogen,is recycled). Examples of suitable spray dryers for use in FSD includedryers from Niro (e.g., the PSD line of spray driers manufactured byNiro: PHARMASD™; Chemical or SD line dryers). FSD can essentially beperformed in any spray dryer that is configured to allow for there-introduction of fines into the drying chamber.

Additional post drying, e.g., in a vacuum or fluidized bed dryer or adouble cone or biconical post-dryer or a tumble dryer, can be performedif needed/applicable to remove further solvents. In some embodiments, apost-drying step is performed.

To remove the solvent or solvent mixture, vacuum drying, spray drying,fluidized spray drying, tray drying, lyophilization, rotovapping, andother drying procedures may be applied. Applying any of these methodsusing appropriate processing parameters, according to this disclosure,would provide Compound 1, or a pharmaceutically acceptable salt thereofin an amorphous state in the final solid dispersion product. Upon use ofappropriate conditions (e.g., low outlet temperatures in the spraydryer, use of low boiling point solvents, use of heated gas) that resultin a dispersion, e.g., powder, with desirable properties (e.g., medianparticle size (d50) of 40-200 microns 9 e.g., 40-150 microns), powderbulk density of >0.2 g/ml (e.g., 0.2 to 0.5 g/ml), or >0.25 g/ml,improved powder flowability (e.g., low cohesion forces, lowinterparticle internal friction); and/or dry powder with low OVIs(Organic Volatile Impurities), e.g., below ICH limits and/or userspecifications), the dispersion can be directly compressed into a dosageform.

In some embodiments, the inlet temperature is between about 50° C. andabout 200° C., e.g., between about 60° C. and about 150° C., betweenabout 70° C. and about 100° C., between about 60° C. and about 95° C.,between about 65° C. and about 85° C., between about 70° C. and about90° C., between about 85° C. and about 95° C., or between about 70° C.and about 85° C.

In some embodiments, the outlet temperature is between about roomtemperature (e.g., USP room temperature (e.g., 21±4° C.)) and about 80°C., e.g., between about 25° C. and about 75° C., between about 30° C.and about 65° C., between about 35° C. and about 70° C., between about40° C. and about 65° C., between about 45° C. and about 60° C., betweenabout 35° C. and about 45° C., between about 35° C. and about 40° C., orbetween about 37° C. and about 40° C.

In some embodiments, the temperature set points of the fluidized beds(the temperature for each bed being selected independently from thetemperature selected for another bed) is between about room temperature(e.g., USP room temperature (e.g., 21±4° C.)) and about 100° C., e.g.,between about 30° C. and about 95° C., between about 40° C. and about90° C., between about 50° C. and about 80° C., between about 60° C. andabout 85° C., between about 65° C. and about 95° C., or between about80° C. and about 95° C.

FSD can be performed on a mixture containing a compound of interest(e.g., a therapeutic agent (e.g., therapeutically active compound),e.g., Compound 1, or a pharmaceutically acceptable salt thereof). Forexample, FSD can be performed on a mixture containing Compound 1, or apharmaceutically acceptable salt thereof (e.g., and one or morepolymer(s), and optionally one or more surfactant(s), and optionally oneor more additional excipients(s)) to obtain a solid dispersion ofamorphous Compound 1, or a pharmaceutically acceptable salt thereof,e.g., that can be directly compressed into an oral dosage form (e.g.,tablet).

Alternatively, the dispersion can be blended with one or more excipientsprior to compression.

In one embodiment, the process for preparing a solid dispersion ofCompound 1 comprises:

a) forming a mixture of Compound 1, or a pharmaceutically acceptablesalt thereof, one or more polymer(s), and one or more solvent(s); and

b) rapidly removing the solvent(s) from the solution to form a solidamorphous dispersion comprising Compound 1, or a pharmaceuticallyacceptable salt thereof, and the one or more polymer(s). The one or morepolymer(s) and one or more solvent(s) may be any of those disclosedherein.

In some embodiments, the solvent is removed by spray drying. In someembodiments the solid dispersion is tray dried using a convection traydryer. In some embodiments, the solid dispersion is screened.

In one embodiment, Compound 1, or a pharmaceutically acceptable saltthereof, is crystalline. In another embodiment, Compound 1, or apharmaceutically acceptable salt thereof, is amorphous.

As would be appreciated by one of skill in the art, spray drying may bedone and is often done in the presence of an inert gas such as nitrogen.In certain embodiments, processes that involve spray drying may be donein the presence of a supercritical fluid involving carbon dioxide or amixture including carbon dioxide.

In another embodiment, the process for preparing a solid dispersion ofCompound 1, or a pharmaceutically acceptable salt thereof, comprises:

a) forming a mixture of Compound 1, or a pharmaceutically acceptablesalt thereof, a polymer, and a solvent; and

b) spray-drying the mixture to form a solid dispersion comprisingCompound 1, or a pharmaceutically acceptable salt thereof, and thepolymer.

Post-drying and/or polishing the wet spray dried dispersion to below ICHor given specifications for residual solvents can optionally beperformed.

These processes may be used to prepare the pharmaceutical compositionsdisclosed herein. The amounts and the features of the components used inthe processes may be as disclosed herein.

In some embodiments, the solvent comprises one or more volatilesolvent(s) to dissolve or suspend Compound 1, or a pharmaceuticallyacceptable salt thereof, and the polymer(s). In some embodiments, theone or more solvent(s) completely dissolves Compound 1, or apharmaceutically acceptable salt thereof, and the polymer(s).

In some embodiments, the one or more solvent(s) is a volatile solvent(e.g., methylene chloride, acetone, methanol, ethanol, chloroform,tetrahydrofuran (THF), or a mixture thereof). Examples of suitablevolatile solvents include those that dissolve or suspend thetherapeutically active compound either alone or in combination withanother co-solvent. In some embodiments, the solvent(s) completelydissolves the therapeutically active compound. In some embodiments, thesolvent is acetone. In some embodiments, the solvent is methanol.

In some embodiments, the solvent is a non-volatile solvent (e.g.,organic acids such as glacial acetic acid, dimethyl sulfoxide (DMSO),dimethylformamide (DMF), or water). In some embodiments, a non-volatilesolvent is a component in a solvent system. For example the non-volatilesolvent is present as a component in a solvent from about 1% to about20% w/w (e.g., from about 3% w/w to about 15% w/w, from about 4% w/w toabout 12% w/w, or from about 5% w/w to about 10% w/w).

In some embodiments, the solvent is a mixture of solvents. For example,the solvent can include from about 0% to about 30% acetone and fromabout 70% to about 100% methanol, or the solvent can include from about0% to about 40% acetone and from about 60% to about 100% methanol. Otherexemplary ratios of methanol to acetone include 80:20, 75:25, 70:30,60:40, 55:45, and 50:50.

In some embodiments, the solvent is a combination of solvents includingat least one non-volatile solvent. For example, the solvent is acombination of components that includes both a volatile solvent and anon-volatile solvent. In some embodiments, the solvent system is acombination of a volatile solvent or combination of solvents such asmethanol and acetone with a non-volatile solvent such as glacial aceticacid. For example, the solvent system comprises from about 40% to about80% methanol, from about 20% to about 35% acetone, and from about 1% toabout 15% glacial acetic acid (e.g., from about 50% to about 70%methanol, from about 25% to about 30% acetone, and from about 3% toabout 12% glacial acetic acid).

In some embodiments, the solvent system is a combination of a volatilesolvent or combination of solvents such as methanol and acetone with anon-volatile solvent such as water. For example, the solvent systemcomprises from about 40% to about 80% methanol, from about 20% to about35% acetone, and from about 0.1% to about 15% water (e.g., from about50% to about 70% methanol, from about 25% to about 30% acetone, and fromabout 1% to about 5% water).

Pharmaceutical Compositions

Pharmaceutical compositions of the solid dispersion may be made by aprocess described herein. For example, a solid dispersion of: (a)Compound 1, or a pharmaceutically acceptable salt thereof, and (b) oneor more polymer(s), and optionally one or more surfactant(s) andoptionally one or more additional excipient(s).

Provided herein are pharmaceutical compositions, comprising: (a) a soliddispersion, comprising Compound 1, or a pharmaceutically acceptable saltthereof, and a polymer; and (b) one or more pharmaceutically acceptablecarrier(s). Examples of pharmaceutically acceptable carriers arefillers, disintegrants, wetting agents, glidants, and lubricants.

In some embodiments, the pharmaceutical compositions may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, emulsions and aqueous suspensions,dispersions and solutions.

In some embodiments the pharmaceutical composition is a tablet.

In some embodiments the pharmaceutical composition comprises a directlycompressed dosage form of Compound 1, or a pharmaceutically acceptablesalt thereof.

In some embodiments, the pharmaceutical composition also includes afiller. The filler can be, for example, microcrystalline cellulose,lactose, mannitol, ethyl cellulose, sorbitol, starch, sucrose, calciumphosphate, powdered cellulose, silicified microcrystalline cellulose,isomalt, or mixtures thereof. In some embodiments, the filler ismicrocrystalline cellulose.

In some embodiments, the filler is present in the pharmaceuticalcomposition in an amount of between about 10% w/w and 50% w/w (e.g.,between about 15% w/w and about 45% w/w; between about 20% w/w and about40% w/w; between about 25% w/w and about 35% w/w; or between about 28%w/w and about 32% w/w). In some embodiments, the filler is present inthe pharmaceutical composition in an amount of from about 20% w/w toabout 35% w/w, for example from about 25% w/w to about 34% w/w, or fromabout 26% w/w to about 33% w/w, or from about 27% w/w to about 32% w/w,for example, about 28% w/w, about 28.5% w/w, about 29% w/w, about 29.5%w/w about 30% w/w, about 30.5% w/w, about 31% w/w, or about 31.5% w/w.In some embodiments, the filler is present in the pharmaceuticalcomposition in an amount of about 29% w/w, about 29.1% w/w, about 29.2%w/w, about 29.3% w/w, about 29.4% w/w, about 29.5% w/w, about 29.6% w/w,about 29.7% w/w, about 29.8% w/w, about 29.9% w/w, or about 30% w/w. Insome embodiments, the filler is present in the pharmaceuticalcomposition in an amount of between about 25% w/w and about 35% w/w. Insome embodiments, the filler is present in the pharmaceuticalcomposition in an amount of about 29.5% w/w.

In some embodiments, the pharmaceutical composition also includes adisintegrant. The disintegrant can be, for example, colloidal silicondioxide, powdered cellulose, calcium silicate, crospovidone, calciumalginate, methyl cellulose, chitosan, carboxy methyl cellulose,croscarmellose sodium, carboxymethyl starch, sodium alginate, sodiumstarch glycolate, pregelatinized starch, or mixtures thereof. In someembodiments, the disintegrant is croscarmellose sodium.

In some embodiments, the disintegrant is present in the pharmaceuticalcomposition in an amount of between about 1% w/w and 15% w/w (e.g.,between about 3% w/w and about 12% w/w; between about 4% w/w and about10% w/w; between about 5% w/w and about 7% w/w; or between about 6% w/wand about 7% w/w). In some embodiments, the disintegrant is present inthe pharmaceutical composition in an amount of about 3% w/w, about 3.5%w/w, about 4% w/w, about 49.5% w/w about 5% w/w, about 5.5% w/w, about6% w/w, or about 6.5% w/w, about 7% w/w, about 7.5% w/w, about 8% w/w,about 8.5% w/w, about 9% w/w, about 9.5% w/w, or about 10% w/w. In someembodiments, the disintegrant is present in the pharmaceuticalcomposition in an amount of between about 5% w/w and about 7% w/w. Insome embodiments, the disintegrant is present in the pharmaceuticalcomposition in an amount of about 6% w/w.

In some embodiments, the pharmaceutical composition also includes awetting agent. The wetting agent can be, for example, sodium laurylsulfate, sodium dodecyl sulfate, polysorbates (such as Tween 20 andTween 80), poloxamers (such as Poloxamer 335 and Poloxamer 407),glyceryl monooleate, or mixtures thereof. In some embodiments, thewetting agent is sodium lauryl sulfate.

In some embodiments, the wetting agent is present in the pharmaceuticalcomposition in an amount of between about 0.1% w/w and 2% w/w (e.g.,between about 0.5% w/w and about 2% w/w; between about 0.5% w/w andabout 1.5% w/w; or between about 1% w/w and about 1.5% w/w). In someembodiments, the wetting agent is present in the pharmaceuticalcomposition in an amount of about 0.1% w/w, about 0.2% w/w, about 0.3%w/w, about 0.4% w/w about 0.5% w/w, about 0.6% w/w, about 0.7% w/w, orabout 0.8% w/w, about 0.9% w/w, about 1% w/w, about 1.1% w/w, about 1.2%w/w, about 1.3% w/w, about 1.4% w/w, about 1.5% w/w, about 1.6% w/w,about 1.7% w/w, about 1.8% w/w, about 1.9% w/w, or about 2% w/w. In someembodiments, the wetting agent is present in the pharmaceuticalcomposition in an amount of between about 0.5% w/w and about 1.5% w/w.In some embodiments, the wetting agent is present in the pharmaceuticalcomposition in an amount of about 1% w/w.

In some embodiments, the pharmaceutical composition also includes aglidant. The glidant can be, for example, silicon dioxide, colloidalsilicon dioxide, tribasic calcium phosphate, magnesium stearate,magnesium trisilicate, powdered cellulose, talc, starch, and mixturesthereof. In some embodiments, the glidant is colloidal silicon dioxide.

In some embodiments, the glidant is present in the pharmaceuticalcomposition in an amount of between about 0.1% w/w and 5% w/w (e.g.,between about 1% w/w and about 4% w/w; between about 1% w/w and about 3%w/w; or between about 1.5% w/w and about 2.5% w/w). In some embodiments,the glidant is present in the pharmaceutical composition in an amount ofabout 0.5% w/w, about 1% w/w, about 1.5% w/w, about 2% w/w about 2.5%w/w, about 3% w/w, about 3.5% w/w, or about 4% w/w, about 4.5% w/w, orabout 5% w/w. In some embodiments, the glidant is present in thepharmaceutical composition in an amount of about 1.1% w/w, about 1.2%w/w, about 1.3% w/w, about 1.4% w/w, about 1.5% w/w, about 1.6% w/w,about 1.7% w/w, about 1.8% w/w, about 1.9% w/w, about 2% w/w, 2.1% w/w,about 2.2% w/w, about 2.3% w/w, about 2.4% w/w, about 2.5% w/w, about2.6% w/w, about 2.7% w/w, about 2.8% w/w, about 2.9% w/w, or about 3%w/w. In some embodiments, the glidant is present in the pharmaceuticalcomposition in an amount of between about 1% w/w and about 3% w/w. Insome embodiments, the glidant is present in the pharmaceuticalcomposition in an amount of about 2% w/w.

In some embodiments, the pharmaceutical composition also includes alubricant. The lubricant can be, for example, magnesium stearate, talc,sodium stearyl fumarate, glyceryl behenate, hydrogenated vegetable oil,zinc stearate, calcium stearate, sucrose stearate, polyvinyl alcohol,magnesium lauryl sulfate, or mixtures thereof. In some embodiments, thelubricant is magnesium stearate.

In some embodiments, the lubricant is present in the pharmaceuticalcomposition in an amount of between about 0.1% w/w and 5% w/w (e.g.,between about 1% w/w and about 4% w/w; between about 1% w/w and about 3%w/w; or between about 1% w/w and about 2% w/w). In some embodiments, thelubricant is present in the pharmaceutical composition in an amount ofabout 0.5% w/w, about 1% w/w, about 1.5% w/w, about 2% w/w about 2.5%w/w, about 3% w/w, about 3.5% w/w, or about 4% w/w, about 4.5% w/w, orabout 5% w/w. In some embodiments, the lubricant is present in thepharmaceutical composition in an amount of about 0.1% w/w, about 0.2%w/w, about 0.3% w/w, about 0.4% w/w, about 0.5% w/w, about 0.6% w/w,about 0.7% w/w, about 0.8% w/w, about 0.9% w/w, about 1% w/w, about 1.1%w/w, about 1.2% w/w, about 1.3% w/w, about 1.4% w/w, about 1.5% w/w,about 1.6% w/w, about 1.7% w/w, about 1.8% w/w, about 1.9% w/w, about 2%w/w, 2.1% w/w, about 2.2% w/w, about 2.3% w/w, about 2.4% w/w, or about2.5% w/w. In some embodiments, the lubricant is present in thepharmaceutical composition in an amount of between about 0.5% w/w andabout 2.5% w/w. In some embodiments, the lubricant is present in thepharmaceutical composition in an amount of about 1.5% w/w.

In some embodiments, the solid dispersion makes up about 25% to 85% byweight of the total weight of the pharmaceutical composition. In someembodiments, the solid dispersion makes up about 50% to about 70% byweight of the total weight of the pharmaceutical composition.

In some embodiments, the Compound 1, or a pharmaceutically acceptablesalt thereof makes up about 15% to 45% of the total weight of thepharmaceutical composition, and the one or more polymer(s) makes upabout 15% to 45% of the total weight of the pharmaceutical composition.

In some embodiments, the Compound 1, or a pharmaceutically acceptablesalt thereof makes up about 20% w/w of the pharmaceutical composition,the one or more polymer(s) makes up about 40% w/w of the pharmaceuticalcomposition.

In some embodiments, the Compound 1, or a pharmaceutically acceptablesalt thereof makes up about 25% w/w of the pharmaceutical composition,the one or more polymer(s) makes up about 35% w/w of the pharmaceuticalcomposition.

In some embodiments, the Compound 1, or a pharmaceutically acceptablesalt thereof makes up about 30% w/w of the pharmaceutical composition,the one or more polymer(s) makes up about 30% w/w of the pharmaceuticalcomposition.

In some embodiments, the Compound 1, or a pharmaceutically acceptablesalt thereof makes up about 35% w/w of the pharmaceutical composition,the one or more polymer(s) makes up about 25% w/w of the pharmaceuticalcomposition.

In some embodiments, the solid dispersion makes up from between about50% w/w to about 70% w/w of the pharmaceutical composition, the fillermakes up from between about 25% w/w to about 35% w/w of thepharmaceutical composition, the disintegrant makes up from between about5% w/w to about 7% w/w of the pharmaceutical composition, the wettingagent makes up from between about 0.5% w/w to about 1.5% w/w of thepharmaceutical composition, the glidant makes up from between about 1%w/w to about 3% w/w of the pharmaceutical composition, the lubricantmakes up from between about 0.5% w/w to about 2.5% w/w of thepharmaceutical composition thereby totaling 100% by weight of thecomposition.

In some embodiments, the solid dispersion makes up about 60% w/w of thepharmaceutical composition, the filler makes up about 29.5% w/w of thepharmaceutical composition, the disintegrant makes up about 6% w/w ofthe pharmaceutical composition, the wetting agent makes up about 1% w/wof the pharmaceutical composition, the glidant makes up about 2% w/w ofthe pharmaceutical composition, the lubricant makes up about 1.5% w/w ofthe pharmaceutical composition.

In some embodiments, the pharmaceutical composition comprises, frombetween about 25% w/w to about 35% w/w of Compound 1, or apharmaceutically acceptable salt thereof, from between about 25% w/w toabout 35% w/w of hypromellose acetate succinate (HPMCAS), from betweenabout 25% w/w to about 35% w/w of microcrystalline cellulose, frombetween about 5% w/w to about 7% w/w croscarmellose sodium, from betweenabout 0.5% w/w to about 1.5% w/w sodium lauryl sulfate, about frombetween about 1% w/w to about 3% w/w colloidal silicon dioxide, and frombetween about 0.5% w/w to about 2.5% w/w of magnesium stearate, therebytotaling 100% by weight of the composition.

In some embodiments, the pharmaceutical composition comprises, about 30%w/w of Compound 1, or a pharmaceutically acceptable salt thereof, about30% w/w of hypromellose acetate succinate (HPMCAS), about 29.5% w/w ofmicrocrystalline cellulose, about 6% w/w croscarmellose sodium, about 1%w/w sodium lauryl sulfate, about 2% w/w colloidal silicon dioxide, andabout 1.5% w/w of magnesium stearate.

In some embodiments, the solid dispersion, filler, disintegrant, wettingagent, glidant, and lubricant are added intragranularly. In someembodiments, an additional amount of the filler, disintegrant, glidant,and lubricant are added extragranularly.

In some embodiments, the pharmaceutical composition comprises, thefollowing intragranularly added components: the solid dispersion makesup from about 50% w/w to about 70% w/w of the pharmaceuticalcomposition, the filler makes up from about 18% w/w to about 26% w/w ofthe pharmaceutical composition, disintegrant makes up from about 2% w/wto about 6% w/w of the pharmaceutical composition, wetting agent makesup from about 0.5% w/w to about 1.5% w/w of the pharmaceuticalcomposition, glidant makes up from about 0.5% w/w to about 1.5% w/w ofthe pharmaceutical composition, and lubricant makes up from about 0.25%w/w to about 1% w/w of the pharmaceutical composition.

In some embodiments, a the pharmaceutical composition comprises thefollowing extragranularly added components: an additional amount of thefiller makes up from about 4% w/w to about 12% w/w of the pharmaceuticalcomposition, an additional amount of the disintegrant makes up fromabout 1% w/w to about 3% w/w of the pharmaceutical composition, anadditional amount of the glidant makes up from about 0.5% w/w to about1.5% w/w of the pharmaceutical composition, and an additional amount ofthe lubricant makes up from about 0.5% w/w to about 1.5% w/w of thepharmaceutical composition, and are added extragranularly.

In some embodiments, the pharmaceutical composition comprises, thefollowing intragranularly added components: the solid dispersion makesup about 60% w/w of the pharmaceutical composition, the filler makes upabout 21.5% w/w of the pharmaceutical composition, disintegrant makes upabout 4% w/w of the pharmaceutical composition, wetting agent makes upabout 1% w/w of the pharmaceutical composition, glidant makes up about1% w/w of the pharmaceutical composition, and lubricant makes up about0.5% w/w of the pharmaceutical composition.

In some embodiments, a the pharmaceutical composition comprises thefollowing extragranularly added components: an additional amount of thefiller makes up about 8% w/w of the pharmaceutical composition, anadditional amount of the disintegrant makes up about 2% w/w of thepharmaceutical composition, an additional amount of the glidant makes upabout 1% w/w of the pharmaceutical composition, and an additional amountof the lubricant makes up about 1% w/w of the pharmaceuticalcomposition, and are added extragranularly.

In some embodiments, the pharmaceutical composition comprises, thefollowing intragranularly added components: the solid dispersioncomprising Compound 1, or a pharmaceutically acceptable salt thereof,and hypromellose acetate succinate (HPMCAS), makes up from about 50% w/wto about 70% w/w of the pharmaceutical composition, microcrystallinecellulose makes up from about 18% w/w to about 26% w/w of thepharmaceutical composition, croscarmellose sodium makes up from about 2%w/w to about 6% w/w of the pharmaceutical composition, sodium laurylsulfate makes up from about 0.5% w/w to about 1.5% w/w of thepharmaceutical composition, colloidal silicon dioxide makes up fromabout 0.5% w/w to about 1.5% w/w of the pharmaceutical composition, andmagnesium stearate makes up from about 0.25% w/w to about 1% w/w of thepharmaceutical composition.

In some embodiments, a the pharmaceutical composition comprises thefollowing extragranularly added components: an additional amount ofmicrocrystalline cellulose makes up from about 4% w/w to about 12% w/wof the pharmaceutical composition, an additional amount ofcroscarmellose sodium makes up from about 1% w/w to about 3% w/w of thepharmaceutical composition, an additional amount of colloidal silicondioxide makes up from about 0.5% w/w to about 1.5% w/w of thepharmaceutical composition, and an additional amount of magnesiumstearate makes up from about 0.5% w/w to about 1.5% w/w of thepharmaceutical composition, and are added extragranularly.

In some embodiments, the pharmaceutical composition comprises, thefollowing intragranularly added components: the solid dispersioncomprising Compound 1, or a pharmaceutically acceptable salt thereof,and hypromellose acetate succinate (HPMCAS), makes up about 60% w/w ofthe pharmaceutical composition, microcrystalline cellulose makes upabout 21.5% w/w of the pharmaceutical composition, croscarmellose sodiummakes up about 4% w/w of the pharmaceutical composition, sodium laurylsulfate makes up about 1% w/w of the pharmaceutical composition,colloidal silicon dioxide makes up about 1% w/w of the pharmaceuticalcomposition, and magnesium stearate makes up about 0.5% w/w of thepharmaceutical composition.

In some embodiments, a the pharmaceutical composition comprises thefollowing extragranularly added components: an additional amount ofmicrocrystalline cellulose makes up about 8% w/w of the pharmaceuticalcomposition, an additional amount of croscarmellose sodium makes upabout 2% w/w of the pharmaceutical composition, an additional amount ofcolloidal silicon dioxide makes up about 1% w/w of the pharmaceuticalcomposition, and an additional amount of magnesium stearate makes upabout 1% w/w of the pharmaceutical composition, and are addedextragranularly.

A subject may be administered a dose of Compound 1, or apharmaceutically acceptable salt thereof, as described in Example 5.Lower or higher doses than those recited above may be required. Specificdosage and treatment regimens for any particular subject will dependupon a variety of factors, including the activity of the specificcompound, employed, the age, body weight, general health status, sex,diet, time of administration, rate of excretion, drug combination, theseverity and course of the disease, condition or symptoms, the subject'sdisposition to the disease, condition or symptoms, and the judgment ofthe treating physician.

Upon improvement of a subject's condition, a maintenance dose of acompound, composition or combination of one aspect of this invention maybe administered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level. Subjects may, however,require intermittent treatment on a long-term basis upon any recurrenceof disease symptoms.

Methods of Use

The inhibitory activities of Compound 1, and pharmaceutically acceptablesalts thereof provided herein against IDH1 mutants (e.g., IDH1R132H orIDH1R132C) can be tested by methods described in Example A of PCTPublication No. WO 2013/107291 and US Publication No. US 2013/0190249,hereby incorporated by reference in their entirety, or analogousmethods.

Provided is a method for treating an advanced solid tumor, such as acutemyelogenous leukemia (AML), myelodysplastic syndrome (MDS),myeloproliferative neoplasms (MPN), myeloproliferative neoplasms (MPN),chronic myelomonocytic leukemia (CMML), B-acute lymphoblastic leukemias(B-ALL), or lymphoma (e.g., T-cell lymphoma), each characterized by thepresence of a mutant allele of IDH1, comprising administering to asubject in need thereof a pharmaceutical composition comprising: (a)Compound 1, or a pharmaceutically acceptable salt thereof, as part of asolid dispersion, and optionally (b) one or more pharmaceuticallyacceptable carrier(s). In one embodiment, the advanced solid tumor, suchas acute myelogenous leukemia (AML), myelodysplastic syndrome (MDS),myeloproliferative neoplasms (MPN), myeloproliferative neoplasms (MPN),chronic myelomonocytic leukemia (CMML), B-acute lymphoblastic leukemias(B-ALL), or lymphoma (e.g., T-cell lymphoma), to be treated ischaracterized by a mutant allele of IDH1, wherein the IDH1 mutationresults in a new ability of the enzyme to catalyze the NAPH-dependentreduction of α-ketoglutarate to R(−)-2-hydroxyglutarate in a patient. Inone aspect of this embodiment, the mutant IDH1 has an R132X mutation. Inone aspect of this embodiment, the R132X mutation is selected fromR132H, R132C, R132L, R132V, R132S and R132G. In another aspect, theR132X mutation is R132H or R132C. In yet another aspect, the R132Xmutation is R132H.

Advanced solid tumors, such as acute myelogenous leukemia (AML),myelodysplastic syndrome (MDS), myeloproliferative neoplasms (MPN),myeloproliferative neoplasms (MPN), chronic myelomonocytic leukemia(CMML), B-acute lymphoblastic leukemias (B-ALL), or lymphoma (e.g.,T-cell lymphoma), each characterized by the presence of a mutant alleleof IDH1 can be analyzed by sequencing cell samples to determine thepresence and specific nature of (e.g., the changed amino acid presentat) a mutation at amino acid 132 of IDH1.

Without being bound by theory, applicants believe that mutant alleles ofIDH1 wherein the IDH1 mutation results in a new ability of the enzyme tocatalyze the NAPH-dependent reduction of α-ketoglutarate toR(−)-2-hydroxyglutarate, and in particular R132H mutations of IDH1,characterize a subset of all types of cancers, without regard to theircellular nature or location in the body. Thus, the compounds, andmethods of one aspect of this invention are useful to treat advancedsolid tumors, such as acute myelogenous leukemia (AML), myelodysplasticsyndrome (MDS), myeloproliferative neoplasms (MPN), myeloproliferativeneoplasms (MPN), chronic myelomonocytic leukemia (CMML), B-acutelymphoblastic leukemias (B-ALL), or lymphoma (e.g., T-cell lymphoma),each characterized by the presence of a mutant allele of IDH1 impartingsuch activity and in particular an IDH1 R132H or R132C mutation.

In one embodiment, the efficacy of treatment of advanced hematologicmalignancies, such as acute myelogenous leukemia (AML), myelodysplasticsyndrome (MDS), myeloproliferative neoplasms (MPN), myeloproliferativeneoplasms (MPN), chronic myelomonocytic leukemia (CMML), B-acutelymphoblastic leukemias (B-ALL), or lymphoma (e.g., T-cell lymphoma),each characterized by the presence of a mutant allele of IDH1 ismonitored by measuring the levels of 2HG in the subject. Typicallylevels of 2HG are measured prior to treatment, wherein an elevated levelis indicated for the use of Compound 1, or a pharmaceutically acceptablesalt thereof, to treat the advanced hematologic malignancies, such asacute myelogenous leukemia (AML), myelodysplastic syndrome (MDS),myeloproliferative neoplasms (MPN), myeloproliferative neoplasms (MPN),chronic myelomonocytic leukemia (CMML), B-acute lymphoblastic leukemias(B-ALL), or lymphoma (e.g., T-cell lymphoma), each characterized by thepresence of a mutant allele of IDH1. Once the elevated levels areestablished, the level of 2HG is determined during the course of and/orfollowing termination of treatment to establish efficacy. In certainembodiments, the level of 2HG is only determined during the course ofand/or following termination of treatment. A reduction of 2HG levelsduring the course of treatment and following treatment is indicative ofefficacy. Similarly, a determination that 2HG levels are not elevatedduring the course of or following treatment is also indicative ofefficacy. Typically, the these 2HG measurements will be utilizedtogether with other well-known determinations of efficacy of cancertreatment, such as reduction in number and size of tumors and/or othercancer-associated lesions, evaluation of bone marrow biopsies and/oraspirates, complete blood counts and examination of peripheral bloodfilms, improvement in the general health of the subject, and alterationsin other biomarkers that are associated with cancer treatment efficacy.

2HG can be detected in a sample by the methods of PCT Publication No. WOWO/2011/050210 and US Publication No. US2012/0121515 hereby incorporatedby reference in their entirety, or by analogous methods.

Methods of Evaluating Samples and/or Subjects

This section provides methods of obtaining and analyzing samples and ofanalyzing subjects.

Embodiments of the method comprise evaluation of one or more parametersrelated to IDH1, an alpha hydroxy neoactivity, e.g., 2HG neoactivity,e.g., to evaluate the IDH1 2HG neoactivity genotype or phenotype. Theevaluation can be performed, e.g., to select, diagnose or prognose thesubject, to select a therapeutic agent, e.g., an inhibitor, or toevaluate response to the treatment or progression of disease. In anembodiment the evaluation, which can be performed before and/or aftertreatment has begun, is based, at least in part, on analysis of a tumorsample, cancer cell sample, or precancerous cell sample, from thesubject. E.g., a sample from the patient can be analyzed for thepresence or level of an alpha hydroxy neoactivity product, e.g., 2HG,e.g., R-2HG, by evaluating a parameter correlated to the presence orlevel of an alpha hydroxy neoactivity product, e.g., 2HG, e.g., R-2HG.An alpha hydroxy neoactivity product, e.g., 2HG, e.g., R-2HG, in thesample can be determined by a chromatographic method, e.g., by LC-MSanalysis. It can also be determined by contact with a specific bindingagent, e.g., an antibody, which binds the alpha hydroxy neoactivityproduct, e.g., 2HG, e.g., R-2HG, and allows detection. In an embodimentthe sample is analyzed for the level of neoactivity, e.g., an alphahydroxy neoactivity, e.g., 2HG neoactivity. In an embodiment the sampleis analysed for the presence of a mutant IDH1, protein having an alphahydroxy neoactivity, e.g., 2HG neoactivity (or a corresponding RNA).E.g., a mutant protein specific reagent, e.g., an antibody thatspecifically binds an IDH1 mutant protein, e.g., an antibody thatspecifically binds an IDH1-R132H mutant protein, can be used to detectneoactive mutant enzyme In an embodiment a nucleic acid from the sampleis sequenced to determine if a selected allele or mutation of IDH1disclosed herein is present. In an embodiment the analysis is other thandirectly determining the presence of a mutant IDH1 protein (orcorresponding RNA) or sequencing of an IDH1 gene. In an embodiment theanalysis is other than directly determining, e.g., it is other thansequencing genomic DNA or cDNA, the presence of a mutation at residue132 of IDH1. E.g., the analysis can be the detection of an alpha hydroxyneoactivity product, e.g., 2HG, e.g., R-2HG, or the measurement of themutation's an alpha hydroxy neoactivity, e.g., 2HG neoactivity. In anembodiment the sample is removed from the patient and analyzed. In anembodiment the evaluation can include one or more of performing theanalysis of the sample, requesting analysis of the sample, requestingresults from analysis of the sample, or receiving the results fromanalysis of the sample. (Generally herein, analysis can include one orboth of performing the underlying method or receiving data from anotherwho has performed the underlying method.)

In an embodiment the evaluation, which can be performed before and/orafter treatment has begun, is based, at least in part, on analysis of atissue (e.g., a tissue other than a tumor sample), or bodily fluid, orbodily product. Exemplary tissues include lymph node, skin, hairfollicles and nails. Exemplary bodily fluids include blood, plasma,urine, lymph, tears, sweat, saliva, semen, and cerebrospinal fluid.Exemplary bodily products include exhaled breath. E.g., the tissue,fluid or product can be analyzed for the presence or level of an alphahydroxy neoactivity product, e.g., 2HG, e.g., R-2HG, by evaluating aparameter correlated to the presence or level of an alpha hydroxyneoactivity product, e.g., 2HG, e.g., R-2HG. An alpha hydroxyneoactivity product, e.g., 2HG, e.g., R-2HG, in the sample can bedetermined by a chromatographic method, e.g., by LC-MS analysis. It canalso be determined by contact with a specific binding agent, e.g., anantibody, which binds the alpha hydroxy neoactivity product, e.g., 2HG,e.g., R-2HG, and allows detection. In embodiments where sufficientlevels are present, the tissue, fluid or product can be analyzed for thelevel of neoactivity, e.g., an alpha hydroxy neoactivity, e.g., the 2HGneoactivity. In an embodiment the sample is analysed for the presence ofa mutant IDH1 protein having an alpha hydroxy neoactivity, e.g., 2HGneoactivity (or a corresponding RNA). E.g., a mutant protein specificreagent, e.g., an antibody that specifically binds an IDH mutantprotein, e.g., an antibody that specifically binds an IDH1-R132H mutantprotein can be used to detect neoactive mutant enzyme. In an embodimenta nucleic acid from the sample is sequenced to determine if a selectedallele or mutation of IDH1 disclosed herein is present. In an embodimentthe analysis is other than directly determining the presence of a mutantIDH1 protein (or corresponding RNA) or sequencing of an IDH1 gene. E.g.,the analysis can be the detection of an alpha hydroxy neoactivityproduct, e.g., 2HG, e.g., R-2HG, or the measurement of 2HG neoactivity.In an embodiment the tissue, fluid or product is removed from thepatient and analyzed. In an embodiment the evaluation can include one ormore of performing the analysis of the tissue, fluid or product,requesting analysis of the tissue, fluid or product, requesting resultsfrom analysis of the tissue, fluid or product, or receiving the resultsfrom analysis of the tissue, fluid or product.

In an embodiment the evaluation, which can be performed before and/orafter treatment has begun, is based, at least in part, on alpha hydroxyneoactivity product, e.g., 2HG, e.g., R-2HG, imaging of the subject. Inembodiments magnetic resonance methods are is used to evaluate thepresence, distribution, or level of an alpha hydroxy neoactivityproduct, e.g., 2HG, e.g., R-2HG, in the subject. In an embodiment thesubject is subjected to imaging and/or spectroscopic analysis, e.g.,magnetic resonance-based analysis, e.g., MRI and/or MRS e.g., analysis,and optionally an image corresponding to the presence, distribution, orlevel of an alpha hydroxy neoactivity product, e.g., 2HG, e.g., R-2HG,or of the tumor, is formed. Optionally the image or a value related tothe image is stored in a tangible medium and/or transmitted to a secondsite. In an embodiment the evaluation can include one or more ofperforming imaging analysis, requesting imaging analysis, requestingresults from imaging analysis, or receiving the results from imaginganalysis.

In one embodiment 2HG is directly evaluated.

In another embodiment a derivative of 2HG formed in process ofperforming the analytic method is evaluated. By way of example such aderivative can be a derivative formed in MS analysis. Derivatives caninclude a salt adduct, e.g., a Na adduct, a hydration variant, or ahydration variant which is also a salt adduct, e.g., a Na adduct, e.g.,as formed in MS analysis.

In another embodiment a metabolic derivative of 2HG is evaluated.Examples include species that build up or are elevated, or reduced, as aresult of the presence of 2HG, such as glutarate or glutamate that willbe correlated to 2HG, e.g., R-2HG.

Exemplary 2HG derivatives include dehydrated derivatives such as thecompounds provided below or a salt adduct thereof:

In one embodiment the advanced hematologic malignancy such as acutemyelogenous leukemia (AML), myelodysplastic syndrome (MDS),myeloproliferative neoplasms (MPN), chronic myelomonocytic leukemia(CMML), B-acute lymphoblastic leukemias (B-ALL), or lymphoma (e.g.,T-cell lymphoma) is a tumor wherein at least 30, 40, 50, 60, 70, 80 or90% of the tumor cells carry an IDH1 mutation, and in particular an IDH1R132H or R132C mutation, at the time of diagnosis or treatment.

In one embodiment, the advanced hematologic malignancy to be treated isAML, characterized by the presence of a mutant allele of IDH1. In someembodiments, the AML is relapsed and/or primary refractory. In someembodiments, the AML is relapsed. In some embodiments, the AML isprimary refractory. In other embodiments, the AML is untreated.

In another embodiment, the advanced hematologic malignancy to be treatedis MDS, characterized by the presence of a mutant allele of IDH1. Inanother embodiment, the advanced hematologic malignancy to be treated isMDS with refractory anemia with excess blasts (subtype RAEB-1 orRAEB-2). In other embodiments, the MDS is considered high-risk by theIPSS-R (Greenberg et al. Blood. 2012; 120(12):2454-65). In otherembodiments, the MDS is recurrent. In other embodiments, the MDS isrefractory. In other embodiments, the subject having MDS is intolerantto established therapy known to provide clinical benefit for theirconditions, according to the treating physician.

In another embodiment, the advanced hematologic malignancy to be treatedis CMML, characterized by the presence of a mutant allele of IDH1. Inanother embodiment, the CMML is relapsed and/or primary refractory. Inanother embodiment, the CMML is relapsed. In another embodiment, theCMML is primary refractory.

Treatment methods described herein can additionally comprise variousevaluation steps prior to and/or following treatment with apharmaceutical composition comprising: (a) Compound 1, or apharmaceutically acceptable salt thereof, as part of a solid dispersion,and optionally (b) one or more pharmaceutically acceptable carrier(s).

In one embodiment, prior to and/or after treatment with a pharmaceuticalcomposition comprising: (a) Compound 1, or a pharmaceutically acceptablesalt thereof, as part of a solid dispersion, and optionally (b) one ormore pharmaceutically acceptable carrier(s), the method furthercomprises evaluating the growth, size, weight, invasiveness, stageand/or other phenotype of the advanced hematologic malignancies, such asacute myelogenous leukemia (AML), myelodysplastic syndrome (MDS),myeloproliferative neoplasms (MPN), chronic myelomonocytic leukemia(CMML), B-acute lymphoblastic leukemias (B-ALL), or lymphoma (e.g.,T-cell lymphoma), each characterized by the presence of a mutant alleleof IDH1.

In one embodiment, prior to and/or after treatment with a pharmaceuticalcomposition comprising: (a) Compound 1, or a pharmaceutically acceptablesalt thereof, as part of a solid dispersion, and optionally (b) one ormore pharmaceutically acceptable carrier(s), the method furthercomprises evaluating the IDH1 genotype of the advanced hematologicmalignancies, such as acute myelogenous leukemia (AML), myelodysplasticsyndrome (MDS), myeloproliferative neoplasms (MPN), myeloproliferativeneoplasms (MPN), chronic myelomonocytic leukemia (CMML), B-acutelymphoblastic leukemias (B-ALL), or lymphoma (e.g., T-cell lymphoma),each characterized by the presence of a mutant allele of IDH1. This maybe achieved by ordinary methods in the art, such as DNA sequencing,immuno analysis, and/or evaluation of the presence, distribution orlevel of 2HG.

In one embodiment, prior to and/or after treatment with a pharmaceuticalcomposition comprising: (a) Compound 1, or a pharmaceutically acceptablesalt thereof, as part of a solid dispersion, and optionally (b) one ormore pharmaceutically acceptable carrier(s), the method furthercomprises determining the 2HG level in the subject. This may be achievedby spectroscopic analysis, e.g., magnetic resonance-based analysis,e.g., MRI and/or MRS measurement, sample analysis of bodily fluid, suchas blood, plasma, urine, or spinal cord fluid analysis, or by analysisof surgical material, e.g., by mass-spectroscopy (e.g. LC-MS, GC-MS), orany of the methods described herein.

Examples General Methods

In the following examples, reagents may be purchased from commercialsources (including Alfa, Acros, Sigma Aldrich, TCI and Shanghai ChemicalReagent Company), and used without further purification.

X-Ray Powder Diffraction (XRPD) parameters: XRPD analysis is performedusing a PANalytical Empyrean X-ray powder diffractometer (XRPD) with a12-auto sample stage. The XRPD parameters used are listed in Table 3.

TABLE 3 Parameters for Reflection Mode Cu, kα, X-Ray wavelength Kα1 (Å):1.540598, Kα2 (Å): 1.544426 Kα2/Kα1 intensity ratio: 0.50 X-Ray tubesetting 45 kV, 40 mA Divergence slit Automatic Scan mode Continuous Scanrange (°2TH) 3°-40° Step size (°2TH) 0.0170 Scan speed (°/min) About 10

Differential Scanning Calorimetry (DSC) parameters: DSC analysis isperformed using a TA Q100, or Q200/Q2000 DSC from TA Instruments. Thetemperature is ramped from room temperature to the desired temperatureat a heating rate of 10° C./min using N₂ as the purge gas, with pancrimped.

Thermogravimetric Analysis (TGA) parameters: TGA analysis is performedusing a TA Q500/Q5000 TGA from TA Instruments. The temperature is rampedfrom room temperature to the desired temperature at a heating rate of10° C./min or 20° C./min using N₂ as the purge gas.

Example 1

Compound 1 and various amounts of Hypromellose Acetate Succinate-MG(Hypromellose Acetate Succinate, MG grade, Shin-Etsu Chemical Co.)polymer may be used to generate the amorphous solid dispersionintermediate and formulation presented in this Example 1. Successcriteria may include manufacturing the batches with reasonable yield(>60%), low residual solvents (≤3000 ppm), as well as meetingspecifications for assay and purity.

Step 1: Preparation of Compound 1 Amorphous Solid Dispersion

Form 1 and hypromellose acetate succinate (HPMCAS) (50%/50%, w/w) areweighed and dissolved in methanol and spray-dried (Buchi B-290) toproduce an amorphous Compound 1 and hypromellose acetate succinate(HPMCAS) solid dispersion. Spray drying processing parameters includenitrogen as the drying gas, an inlet temperature of about 85° C. to 95°C., an outlet temperature of about 37° C. to 40° C., spray solutionconcentration of about 5% w/w/, secondary drying of 12 to 18 hours at40° C. The amorphous solid dispersion is further dried in a vacuum ovenand then screened. The amorphous solid dispersion may be packaged indouble polyethylene bags with twisted nylon tie and placed in a highdensity polyethylene (HDPE) container containing desiccant and stored at2-8° C. until the next step of processing.

Step 2: Manufacture of Compound 1 Tablets

Compound 1 and hypromellose acetate succinate amorphous solid dispersionintermediate and all other excipients disclosed in Table 4 are weighedand sieved for blending.

Weighing and Screening Intragranular Ingredients

Compound 1 and hypromellose acetate succinate amorphous solid dispersionis mixed with microcrystalline cellulose, croscarmellose sodium, sodiumlauryl sulfate, colloidal silicon dioxide, and magnesium stearate in asuitable blender.

TABLE 4 Batch formulation composition Amount per batch (g) 50 mg 200 mgComponent Function tablet tablet Intragranular Compound 1*Therapeutically 241.75 1204.01 Active Compound Hypromellose Stabilizer241.75 1204.01 Acetate Succinate* Microcrystalline Filler 173.26 862.87Cellulose Croscarmellose Disintegrant 32.23 160.53 Sodium Sodium LaurylWetting 8.06 40.13 Sulfate agent Colloidal Silicon Glidant 8.06 40.13Dioxide Magnesium Stearate Lubricant 4.03 20.07 ExtragranularMicrocrystalline Filler 64.47 321.07 Cellulose CroscarmelloseDisintegrant 16.12 80.27 Sodium Colloidal Silicon Glidant 8.06 40.13Dioxide Magnesium Stearate Lubricant 8.06 40.13 Total 805.85 4013.36Theoretical number of tablets 4835 6020 *Compound 1 and HypromelloseAcetate Succinate amorphous solid dispersion intermediate

Intragranule Blending

The intra-granule blend is roller compacted and the compacted materialis sized to produce granules.

Dry Granulation/Sizing

Extra-granular microcrystalline cellulose, croscarmellose sodium,colloidal silicon and magnesium stearate are weighed and sieved forblending.

Weighing and Screening Extragranular Ingredients

The screened granules and extra-granular excipients are added to asuitable blender and blended.

Extragranule Blending

The blend is compressed using a rotary tablet press set-up tomanufacture tablets of the appropriate shape/size and required weight,thickness, and hardness.

Compression

Bulk Compound 1 tablets are packaged in double sealed polyethylene bagscontaining 30 g silica gel packs which are placed in foil lined drumsand stored at 2-8° C. Tablets are subsequently packaged.

TABLE 5 Tablet composition 50 mg Tablet 200 mg Tablet Amount per ContentAmount per Content Component Function Tablet (mg) (%) Tablet (mg) (%)Compound 1* Therapeutically 50.0 30 200.0 30 Active CompoundHypromellose Stabilizer 50.0 30 200.0 30 Acetate Succinate*Microcrystalline Filler 49.2 29.5 196.7 29.5 Cellulose CroscarmelloseDisintegrant 10.0 6 40.0 6 Sodium Sodium Lauryl Wetting 1.7 1 6.8 1Sulfate agent Colloidal Silicon Glidant 3.3 2 13.2 2 Dioxide MagnesiumLubricant 2.5 1.5 10.0 1.5 Stearate Total 166.7 100.0 666.7 100.0

Example 2 Synthesis of Form 1

A mixture of Compound 1 (3.5 kg, 7.28 mol) in 1,4-dioxane (35 L) isdegassed by N₂ bubbling for a maximum of 20 min.2-chloro-4-cyanopyridine (1.21 kg, 8.73 mol),tris(dibenzylideneacetone)-dipalladium(0) (167 g, 0.18 mol), and4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (xantphos) (211 g, 0.36mol) are added and the reaction mixture is degassed by N₂ bubbling for amaximum of 10 min. K2CO₃ (1.21 kg, 8.73 mol) is added and the reactionmixture is degassed by N₂ bubbling for a maximum of 30 min. The reactionmixture is heated at 90-100° C. for 4 to 24 hours until the reaction iscomplete. The reaction mixture is then cooled to 15-25° C. and filteredthrough Celite and is washed with ethyl acetate, and the combinedfiltrate and wash are concentrated.

The 1,4-dioxane is removed, and the residual solid is dissolved in ethylacetate (77.5 L). The ethyl acetate solution is washed successively witha 5% aqueous solution of NaHSO₃, a 2% aqueous solution of EDTA disodium,and a 1% aqueous solution of EDTA disodium salt. The organic phase istreated with activated carbon at 55-65° C. for a maximum of 2 h, and ispurified by silica gel chromatography. After chromatography, theresulting product is purified by two recrystallizations: first Compound1 is dissolved in ethyl acetate and heated to 60-70° C. and heptane isadded. The reaction mixture is cooled to 15-25° C. and stirred for 1-3h. The product is filtered and is dissolved in dichloromethane, then isfiltered and is precipitated with heptane, is filtered and dried toproduce Form 1.

Example 3 Synthesis of Form 2

Method A:

About 100 mg of Compound 1 is mixed with 0.4 mL MeOH and stirred at roomtemperature for 12 h. The suspension is subsequently centrifuged, andthe white solid is isolated.

Method B:

About 10 mg of Compound 1 in 0.2-0.4 mL of a mixture of MeOH:H₂O (9:1)in a 3-mL glass vial. The resulting visually clear solution is coveredwith a cap and subjected to slow evaporation to induce precipitation.The solid is isolated.

Method C:

About 15 mg of Compound 1 is dissolved in a mixture of EtOH:H₂O (8:7volume/volume) or Methyl ethyl ketone (MEK) at 50° C. and stirred at 50°C. for 30 min. Then the solution is cooled slowly to 5° C. at 0.1°C./min, and is stirred at 5° C. overnight. The solid is isolated.

Example 4

The following three homogenous suspensions of Compound 1 are provided:

Form 2 in vehicle (1% d-alpha-tocopheyl polyethylene glycol 1000succinate (TPGS):1% HPMCAS in water), an amorphous solid dispersion of25% w/w Form 2 and 75% w/w HPMCAS-M (Solid Dispersion A) in vehicle, andan amorphous solid dispersion of 25% w/w Form 2 and 75% w/w PVAP (SolidDispersion B) in vehicle (200 mg/kg in 10 mL/kg).

Each suspension is prepared on the day of dosing, and the Sprague Dawleyrats are dosed orally. Serial plasma samples are taken at different timepoints following dosing. Compound 1 concentration in plasma isdetermined using a sensitive and specific LC/MS method. PK parameters,including AUC_(0-72 hr) and Cmax, are calculated using WinNonlinsoftware.

For Form 2, the C_(max) is 1600 ng/mL, and AUC_(0-72 hr) is 21700hr*ng/mL. For solid dispersion A, the C_(max)=6820 ng/mL, andAUC_(0-72 hr) is 105635 hr*ng/mL. For solid dispersion B, the C_(max) is30467 ng/mL; AUC_(0-72 hr) is 406841 hr*ng/mL.

The AUC_(0-72 hr) ratio of Solid Dispersion B to Form 2 is 19. TheAUC_(0-72 hr) ratio of Solid Dispersion A to Form 2 is 5.

Example 5. Phase 1 Clinical Trial Protocol

The safety, PK/PD, and clinical activity evaluation of Compound 1, or apharmaceutically acceptable salt thereof, is evaluated in subjects withadvanced hematologic malignancies, such as AML, MDS, MPN, or CMML), thatharbor an IDH1 mutation. Primary study objectives include 1) assessmentof the safety and tolerability of treatment with Compound 1, or apharmaceutically acceptable salt thereof when administered continuouslyas a single agent dosed orally twice daily (approximately every 12hours) on Days 1 to 28 of a 28-day cycle, and 2) determination of themaximum tolerated dose (MTD) and/or the recommended Phase 2 dose ofCompound 1, or a pharmaceutically acceptable salt thereof in subjects.

Secondary study objectives include 1) description of the dose-limitingtoxicities (DLTs) of Compound 1, or a pharmaceutically acceptable saltthereof in subjects with advanced hematologic malignancies, such as AML,MDS, MPN, or CMML, that harbor an IDH1 mutation, characterization of thepharmacokinetics (PK) of Compound 1, or a pharmaceutically acceptablesalt thereof in subjects with advanced hematologic malignancies, such asAML, MDS, MPN, or CMML, that harbor an IDH1 mutation, 3) evaluation ofthe PK/pharmacodynamic (PD) relationship of Compound 1, or apharmaceutically acceptable salt thereof, and 2-hydroxygluturate (2HG),and 4) characterization of the clinical activity associated withCompound 1, or a pharmaceutically acceptable salt thereof, in subjectswith advanced hematologic malignancies, such as AML, MDS, MPN, or CMML,that harbor an IDH1 mutation.

Exploratory study objectives include 1) evaluation of changes in Ki67levels in tumor samples, 2) characterization of the PD effects ofCompound 1, or a pharmaceutically acceptable salt thereof, in subjectswith advanced hematologic malignancies, such as AML, MDS, MPN, or CMML,that harbor an IDH1 mutation by the assessment of changes in thepatterns of cellular differentiation of isocitrate dehydrogenase-1(IDH1)-mutated tumor cells and changes in histone and deoxyribonucleicacid (DNA) methylation profiles in IDH1-mutated tumor cells, 3)evaluation of gene mutation status, global gene expression profiles, andother potential prognostic markers (cytogenetics) in IDH1-mutated tumorcells, as well as subclonal populations of non-IDH1 mutated tumor cells,to explore predictors of anti-tumor activity and/or resistance, and 4)monitoring plasma cholesterol and 40-OH-cholesterol levels as apotential CYP3A4 induction marker.

Compound 1 will be administered orally twice daily (approximately every12 hours) on Days 1 to 28 in 28-day cycles. If warranted based on theemerging data, an alternative dosing schedule (e.g., once daily or threetimes daily), including administration of the same total daily doseusing different dosing schedules in concurrent cohorts, may be explored.Starting with C1D1, dosing is continuous; there are no inter-cycle restperiods.

Subjects who do not meet any of the standard clinical treatmentwithdrawal criteria may continue treatment beyond Cycle 1.

Subjects will be dispensed the appropriate number of tablets for 28 daysof dosing (plus an additional 2-day supply to allow for scheduling ofvisits) on Day 1 of each cycle. Subjects are to return all unusedtablets (or the empty bottles) on Day 1 of each treatment cycle.Subjects will be given a dosing diary for each treatment cycle. Theyshould record relevant information regarding their study drug in thediary (e.g., confirmation that each daily dose was taken, reasons formissed doses). Treatment compliance will be assessed based on return ofunused drug and the dosing diary.

Subjects should be instructed to take their daily dose at approximatelythe same time each day. Each dose should be taken with a glass of waterand consumed over as short a time as possible. Subjects should beinstructed to swallow tablets whole and to not chew the tablets.Subjects may take Compound 1, or a pharmaceutically acceptable saltthereof with or without food. If the subject forgets to take the dailymorning (or evening) dose, then they should take Compound 1, or apharmaceutically acceptable salt thereof within 6 hours after the misseddose. If more than 6 hours have elapsed, then that dose should beomitted, and the subject should resume treatment with the next scheduleddose.

The study includes a dose escalation phase to determine MTD followed byexpansion cohorts to further evaluate the safety and tolerability of theMTD. The dose escalation phase will utilize a standard “3+3” design.During the dose escalation phase, consented eligible subjects will beenrolled into sequential cohorts of increasing doses of Compound 1, or apharmaceutically acceptable salt thereof. Each dose cohort will plan toenroll a minimum of 3 subjects. The first 3 subjects enrolled in eachdosing cohort during the dose escalation phase of the study willinitially receive a single dose of study drug on Day −3 (i.e., 3 daysprior to the start of daily dosing) and undergo PK/PD assessments over72 hours to evaluate drug concentrations and 2HG levels. The next doseof study drug will be on Cycle 1 Day 1 (C1D1) at which time daily dosingwill begin. The initial dosing regimen will be twice daily(approximately every 12 hours). If warranted based on the emerging data,an alternative dosing schedule (e.g., once daily or three times daily),including administration of the same total daily dose using differentdosing schedules in concurrent cohorts, may be explored. If there aremultiple subjects in the screening process at the time the third subjectwithin a cohort begins treatment, up to 2 additional subjects may beenrolled with approval of the Medical Monitor. For these additionalsubjects, the Day −3 through Day 1 PK/PD assessments are optionalfollowing discussion with the Medical Monitor. The planned doseescalation scheme is illustrated in Table 1.

TABLE 1 Dose Escalation Scheme Compound 1 Number of Cohort Level Dose¹Subjects −1  50 mg² 3 to 6 1 (Starting Dose) 100 mg 3 to 6 2 200 mg 3 to6 3 400 mg 3 to 6 4, etc. 800 mg³ 3 to 6 Expansion MTD⁴ 36⁵ ¹Compound 1,or a pharmaceutically acceptable salt thereof, may be administered twicedaily (approximately every 12 hours). If warranted based on the emergingdata, an alternative dosing schedule (e.g., once daily or three timesdaily), including administration of the same total daily dose usingdifferent dosing schedules in concurrent cohorts, may be explored. ²IfDLTs (are observed at Dose Level 1 (100 mg), the dose for the secondcohort will be decreased to 50 mg (Dose Level −1). ³Continued doublingof the dose until Compound 1-related NCI CTCAE version 4.03 ≥ Grade 2toxicity is observed. Following evaluation of the event(s) by theClinical Study Team, subsequent increases in dose will be guided by theobserved toxicity, and potentially PK and PK/PD data until MTD isdetermined. The absolute percent increase in the dose will be determinedby the Clinical Study Team predicated on the type and severity of anytoxicity seen in the prior dose cohorts. Dose escalation will neverexceed 100%. ⁴Defined as the highest dose that causes DLTs in <1 of 3 or<2 of 6 subjects. If no DLTs are identified, dosing will continue for atleast 2 dose levels above the projected maximum biologically effectiveexposure, as determined by an ongoing assessment of PK/PD and anyobserved clinical activity to determine the recommended Phase 2 dose.⁵To include 3 cohorts of approximately 12 subjects each.

Toxicity severity will be graded according to the National CancerInstitute Common Terminology Criteria for Adverse Events (NCI CTCAE)version 4.03. A DLT is defined as follows. Hematologic includesprolonged myelosuppression, defined as persistence of >3 Gradeneutropenia or thrombocytopenia (by NCI CTCAE, version 4.03,leukemia-specific criteria, i.e., marrow cellularity <5% on Day 28 orlater from the start of study drug without evidence of leukemia) atleast 42 days after the initiation of Cycle 1 therapy. Leukemia-specificgrading should be used for cytopenias (based on percentage decrease frombaseline: 50 to 75%=Grade 3, >75%=Grade 4). All AEs that cannot clearlybe determined to be unrelated to Compound 1, or a pharmaceuticallyacceptable salt thereof will be considered relevant to determining DLTs.

If, after the third subject completes the 28-day DLT evaluation period(i.e., Cycle 1), no DLTs are observed, the study will proceed with doseescalation to the next cohort following safety review by the ClinicalStudy Team. If 1 of 3 subjects experiences a DLT during the first cycle,3 additional subjects will be enrolled in that cohort. If none of theadditional 3 subjects experience a DLT, dose escalation may continue tothe next cohort following safety review. If 2 or more subjects in acohort experience DLTs during the first cycle, dose escalation will behalted and the next lower dose level will be declared the MTD.Alternatively, a dose level intermediate between the dose levelexceeding MTD and the previous does level may be explored and declaredMTD if <2 out of 6 patients experience a DLT at that dose. If the MTDcohort includes only 3 subjects, an additional 3 subjects will beenrolled at that dose level to confirm that <2 of 6 subjects experiencea DLT at that dose.

Increases in the dose of Compound 1, or a pharmaceutically acceptablesalt thereof, for each dose cohort will be guided by an acceleratedtitration design, where the dose will be doubled (100% increase) fromone cohort to the next until Compound 1-related NCI CTCAE version 4.03Grade 2 or greater toxicity is observed in any subject within thecohort. Subsequent increases in dose will be guided by the observedtoxicity, and potentially PK and PK/PD data, until the MTD isdetermined. The absolute percent increase in the daily dose will bedetermined predicated on the type and severity of any toxicity seen inthe prior dose cohorts (but will never exceed 100%). If warranted basedon the emerging data, an alternative dosing schedule (e.g., once dailyor three times daily) may be explored, including administration of thesame total daily dose using different dosing schedules in concurrentcohorts. The MTD is the highest dose that causes DLTs in <2 of 6subjects.

If no DLTs are identified during the dose escalation phase, doseescalation may continue for 2 dose levels above the projected maximumbiologically effective dose, as determined by an ongoing assessment ofPK/PD and any observed clinical activity, to determine the recommendedPhase 2 dose.

To optimize the number of subjects treated at a potentially clinicallyrelevant dose, intra-subject dose escalation will be permitted Followingdetermination of the recommended Phase 2 dose, 3 or more expansioncohorts (with AML, MDS, MPN, or CMML) of approximately 12 subjects eachwill be treated at that dose. The purpose of the expansion cohorts is toevaluate and confirm the safety and tolerability of the recommendedPhase 2 dose in specific disease indications. Subjects enrolled in thesecohorts will undergo the same procedures as subjects in the doseescalation cohorts with the exception that the Day −3 through Day 1PK/PD assessments will be optional.

Subjects will undergo screening procedures within 28 days prior to thestart of study drug treatment to determine eligibility. Screeningprocedures include medical, surgical, and medication history,confirmation of IDH1 mutation via tumor biopsies or leukemic blasts (ifnot documented previously), physical examination, vital signs, EasternCooperative Oncology Group (ECOG) performance status (PS), 12-leadelectrocardiogram (ECG), evaluation of left ventricular ejectionfraction (LVEF), clinical laboratory assessments (hematology, chemistry,coagulation, urinalysis, and serum pregnancy test), bone marrow biopsyand aspirate, and blood and urine samples for 2HG measurement; and bloodsamples for determination of plasma cholesterol and 40-OH-cholesterollevels.

Three days prior to starting the twice daily dosing of Compound 1, or apharmaceutically acceptable salt thereof (Day −3), the first 3 subjectsenrolled in each cohort in the dose escalation phase will receive asingle dose of Compound 1, or a pharmaceutically acceptable salt thereofin clinic and have serial blood and urine samples obtained fordetermination of blood and urine concentrations of Compound 1, or apharmaceutically acceptable salt thereof, its metabolite, and 2HG. Afull 72-hour PK/PD profile will be conducted: subjects will be requiredto remain at the study site for 10 hours on Day −3 and return on Days−2, −1, and 1 for 24, 48, and 72 hour samples, respectively.

Daily treatment with Compound 1, or a pharmaceutically acceptable saltthereof, will begin on C1D1; subjects who did not undergo the Day −3PK/PD assessments will be observed in the clinic for 4 hours followingthe C1D1 dose. The initial dosing regimen will be twice daily(approximately every 12 hours). Safety assessments conducted during thetreatment period include physical examination, vital signs, ECOG PS,12-lead ECGs, LVEF, and clinical laboratory assessments (hematology,chemistry, coagulation, and urinalysis).

All subjects will undergo PK/PD assessments over a 10-hour period onboth C1D15 and C2D1. Additional pre-dose urine and/or blood samplingwill be conducted on C1D8, C1D22, C2D15, C3D1, C3D15, and on Day 1 ofall subsequent cycles. Available bone marrow biopsy samples also will beassessed for 2HG levels.

Subjects will undergo radiographic evaluations (CT/MRI), and assessmentof bone marrow aspirates and biopsies and peripheral blood to assess theextent of disease, at screening, on Day 15, Day 29 and Day 57, and every56 days thereafter while on study drug treatment, independent of dosedelays and/or dose interruptions, and/or at any time when progression ofdisease is suspected. Two core tumor biopsies will be obtained atscreening, at the time of the first assessment of response, and at thetime of disease progression within a window of 3 days around the plannedassessment time point. For patients with acute myelogenous leukemia(AML), response to treatment will be determined by the Investigatorsbased on modified International Working Group (IWG) response criteria.

Subjects may continue treatment with Compound 1, or a pharmaceuticallyacceptable salt thereof until disease progression, occurrence of a DLT,or development of other unacceptable toxicity. All subjects are toundergo an end of treatment assessment (within approximately 5 days ofthe last dose of study drug); in addition, a follow-up assessment is tobe scheduled 28 days after the last dose.

It is estimated that approximately 51 subjects will be enrolled in thestudy. Assuming that identification of the MTD requires the evaluationof 4 dose levels of Compound 1, or a pharmaceutically acceptable saltthereof with only 3 subjects per dose level, with the exception that theMTD requires 6 subjects, then 15 subjects will be enrolled during thedose escalation part of the study. Three cohorts of approximately 12additional subjects each in specific advanced hematologic malignancies(total 36 subjects) will be enrolled in the cohort expansion part of thestudy. Additional subjects may be needed for cohort expansion duringdose escalation, for the replacement of non-evaluable subjects, or forevaluation of alternative dosing regimens other than the plannedescalation scheme or the MTD, to optimize the recommended Phase 2 dose.

A patient must meet all of the following inclusion criteria to beenrolled in the clinical study. 1) Subject must be ≥18 years of age; 2)Subjects must have a) an advanced hematologic malignancy including: i)Relapsed and/or primary refractory AML as defined by World HealthOrganization (WHO) criteria, ii) untreated AML, ≥60 years of age and arenot candidates for standard therapy due to age, performance status,and/or adverse risk factors, according to the treating physician andwith approval of the Medical Monitor, iii) Myelodysplastic syndrome withrefractory anemia with excess blasts (subtype RAEB-1 or RAEB-2), orconsidered high-risk by the Revised International Prognostic ScoringSystem (IPSS-R) (Greenberg et al. Blood. 2012; 120(12):2454-65) that isrecurrent or refractory, or the patient is intolerant to establishedtherapy known to provide clinical benefit for their condition (i.e.,patients must not be candidates for regimens known to provide clinicalbenefit), according to the treating physician and with approval of theMedical Monitor, and iv) Subjects with other relapsed and/or primaryrefractory hematologic cancers, for example CMML, who fulfill theinclusion/excluding criteria may be considered on a case-by case basis;3) subjects must have documented IDH1 gene-mutated disease based onlocal evaluation. Analysis of leukemic blast cells for IDH1 genemutation is to be evaluated at screening (if not evaluated previously)by the site's local laboratory to determine subject eligibility for thestudy. If the site does not have local laboratory access for IDH1 genemutation analysis, central laboratory evaluation is acceptable. Apretreatment tumor sample (from blood and/or bone marrow) will berequired for all screened subjects for central laboratory biomarkeranalysis. Gene mutation analysis of a tumor sample (from blood or bonemarrow) is to be repeated at the End of Treatment visit and submitted tothe central laboratory for biomarker analysis; 4) Subjects must beamenable to serial bone marrow biopsies, peripheral blood sampling, andurine sampling during the study. (The diagnosis and evaluation of AML orMDS can be made by bone marrow aspiration when a core biopsy isunobtainable and/or is not a part of the standard of care. A bone marrowbiopsy is required in case of dry tap or failure (mainly dilution) withthe aspiration.); 5) Subjects or their legal representatives must beable to understand and sign an informed consent; 6) subjects must haveECOG PS of 0 to 2; 7) subjects must have a platelet count ≥20,000/μL(Transfusions to achieve this level are allowed.) Subjects with abaseline platelet count of <20,000/μL due to underlying malignancy areeligible with Medical Monitor approval; 8) Subjects must have adequatehepatic function as evidenced by: a) Serum total bilirubin ≤1.5× upperlimit of normal (ULN), unless considered due to Gilbert's disease orleukemic organ involvement, and b) Aspartate aminotransferase, ALT, andalkaline phosphatase (ALP)≤3.0×ULN, unless considered due to leukemicorgan involvement; 9) Subjects must have adequate renal function asevidenced by a serum creatinine ≤2.0×ULN or Creatinine clearance >40mL/min based on the Cockroft-Gault glomerular filtration rate (GFR)estimation: (140−Age)×(weight in kg)×(0.85 if female)/72× serumcreatinine; 10) Subjects must be recovered from any clinically relevanttoxic effects of any prior surgery, radiotherapy, or other therapyintended for the treatment of cancer. (Subjects with residual Grade 1toxicity, for example Grade 1 peripheral neuropathy or residualalopecia, are allowed with approval of the Medical Monitor.); and 11)Female subjects with reproductive potential must have a negative serumpregnancy test within 7 days prior to the start of therapy. Subjectswith reproductive potential are defined as one who is biologicallycapable of becoming pregnant. Women of childbearing potential as well asfertile men and their partners must agree to abstain from sexualintercourse or to use an effective form of contraception during thestudy and for 90 days (females and males) following the last dose ofCompound 1, or a pharmaceutically acceptable salt thereof.

Compound 1, or a pharmaceutically acceptable salt thereof, will beprovided as 50 and 200 mg strength tablets to be administered orally,twice daily or once daily.

The first 3 subjects in each cohort in the dose escalation portion ofthe study will receive a single dose of study drug on Day −3; their nextdose of study drug will be administered on C1D1 at which time subjectswill start dosing twice daily (approximately every 12 hours) on Days 1to 28 in 28-day cycles. Starting with C1D1, dosing is continuous; thereare no inter-cycle rest periods. Subjects who are not required toundergo the Day −3 PK/PD assessments will initiate twice daily dosing(approximately every 12 hours) with Compound 1, or a pharmaceuticallyacceptable salt thereof on C1D1.

The dose of Compound 1, or a pharmaceutically acceptable salt thereofadministered to a subject will be dependent upon which dose cohort isopen for enrollment when the subject qualifies for the study. Thestarting dose of Compound 1, or a pharmaceutically acceptable saltthereof to be administered to the first cohort of subjects is 100 mgstrength administered orally twice a day (200 mg/day).

Subjects may continue treatment with Compound 1, or a pharmaceuticallyacceptable salt thereof until disease progression, occurrence of a DLT,or development of other unacceptable toxicity.

Criteria for Evaluation

Safety:

AEs, including determination of DLTs, serious adverse events (SAEs), andAEs leading to discontinuation; safety laboratory parameters; physicalexamination findings; vital signs; 12-lead ECGs; LVEF; and ECOG PS willbe monitored during the clinical study. The severity of AEs will beassessed by the NCI CTCAE, Version 4.03.

Compound 1, or a pharmaceutically acceptable salt thereof, may causesensitivity to direct and indirect sunlight. The subjects should bewarned to avoid direct sun exposure. When exposure to sunlight isanticipated for longer than 15 minutes, the subject should be instructedto apply factor 30 or higher sunscreen to exposed areas and wearprotective clothing and sunglasses.

Pharmacokinetics and Pharmacodynamics:

Serial blood samples will be evaluated for determination ofconcentration-time profiles of Compound 1, or a pharmaceuticallyacceptable salt thereof. Urine samples will be evaluated fordetermination of urinary excretion of Compound 1, or a pharmaceuticallyacceptable salt thereof. Blood, bone marrow, and urine samples will beevaluated for determination of 2HG levels. Tumor biopsies will be takenfor evaluation of 2HG and Compound 1, or a pharmaceutically acceptablesalt thereof.

Pharmacokinetic Assessments:

Serial blood samples will be drawn before and after dosing with Compound1, or a pharmaceutically acceptable salt thereof in order to determinecirculating plasma concentrations of Compound 1, or a pharmaceuticallyacceptable salt thereof. The blood samples will also be used for thedetermination of 2HG concentrations and for evaluation of cholesteroland 40-OH-cholesterol levels.

For the first 3 subjects enrolled in a cohort during the dose escalationphase, a single dose of Compound 1, or a pharmaceutically acceptablesalt thereof will be administered on Day −3 (i.e., 3 days prior to theirscheduled C1D1 dose). Blood samples will be drawn prior to thesingle-dose administration of Compound 1, or a pharmaceuticallyacceptable salt thereof and at the following time points afteradministration: 30 minutes and 1, 2, 3, 4, 6, 8, 10, 24, 48, and 72hours. After 72 hours of blood sample collection, subjects will beginoral twice daily dosing of Compound 1, or a pharmaceutically acceptablesalt thereof (i.e., C1D1). The PK/PD profile from Day −3 through Day 1is optional for additional subjects enrolled in the dose escalationphase (i.e., for any subjects beyond the 3 initial subjects enrolled ina cohort) and is not required for subjects enrolled in the expansioncohorts.

All subjects will undergo 10-hour PK/PD sampling on C1D15 and C2D1(i.e., on Days 15 and 29 of twice daily dosing). For this profile, oneblood sample will be drawn immediately prior to that day's first dose ofCompound 1, or a pharmaceutically acceptable salt thereof (i.e., dosingwith Compound 1, or a pharmaceutically acceptable salt thereof willoccur at the clinical site); subsequent blood samples will be drawn atthe following time points after dosing: 30 minutes, and 1, 2, 3, 4, 6,8, and 10 hours. Blood samples also will be drawn on Days 8 and 22 ofCycle 1, Day 15 of Cycle 2, Days 1 and 15 of Cycle 3, and Day 1 of eachcycle thereafter; all samples will be obtained prior to dosing.Additionally, one blood sample will be drawn at the End of TreatmentVisit.

The timing of blood samples drawn for Compound 1, or a pharmaceuticallyacceptable salt thereof concentration determination may be changed ifthe emerging data indicates that an alteration in the sampling scheme isneeded to better characterize the PK profile of Compound 1, or apharmaceutically acceptable salt thereof.

Pharmakodynamic Assessments:

Serial blood samples will be drawn before and after dosing with Compound1, or a pharmaceutically acceptable salt thereof in order to determinecirculating concentrations of 2HG. Samples collected for PK assessmentsalso will be used to assess 2HG levels. In addition, subjects will haveblood drawn for determination of 2HG levels at the screening assessment.

The timing of blood samples drawn for 2HG concentration determinationmay be changed if the emerging data indicate that an alteration in thesampling scheme is needed to better characterize the 2HG response toCompound 1, or a pharmaceutically acceptable salt thereof, treatment.

Urine will be collected for the determination of concentrations of 2HGlevels at the screening assessment and prior to dosing on Day 15 ofCycle 1 and on Day 1 of Cycle 2 and every cycle thereafter. At least 20mL of urine will be collected for each sample.

The volume of each collection will be measured and recorded and sent toa central laboratory for determination of urinary 2HG concentration. Analiquot from each collection will be analyzed for urinary creatinineconcentration.

Tumor biopsy specimens will be collected and assessed for 2HG levels, atthe screening assessment, at the time of the first disease assessment,and at any time disease progression is suspected. A window of 3 daysaround the planned assessment time point is acceptable for all biopsysamples. Tumor biopsies are to be evaluated for morphology and forcellular differentiation via hematoxylin and eosin (H & E) staining andICH for specific cell-type markers. Tumor samples may also be evaluatedfor 2HG levels, Ki67 levels, and, if feasible, intra-tumoral Compound 1,or a pharmaceutically acceptable salt thereof, levels.

Serial blood samples will be drawn to obtain plasma cholesterol and40-OH-cholesterol levels as a potential CYP3A4 induction marker. Samplesare obtained on Day −3 (within 30 minutes), at 24, 48, and 72 hours (±1hour), and on Days 8, 15 and 22 of Cycle 1, Days 1 and 15 of Cycles 2and 3, and Day 1 of every cycle thereafter.

Clinical Activity:

Serial blood and bone marrow biopsies will be evaluated during theclinical study to determine response to Compound 1, or apharmaceutically acceptable salt thereof treatment according to the 2006modified IWG criteria for hematologic malignancies, such as MDS, MDS,MPN or AML (Cheson B D, et al. Blood. 2006; 108(2):419-25).

Disease response to treatment will be assessed through the evaluation ofbone marrow biopsies and/or aspirates, along with complete blood countsand examination of peripheral blood films. Subjects will have the extentof their disease assessed and recorded at screening, on Days 15, 29, and57, every 56 days thereafter while on study drug treatment, independentof dose-delays and/or dose interruptions, and/or at any time whenprogression of disease is suspected. An assessment also will beconducted at the End of Treatment visit for subjects who discontinue thestudy due to reasons other than disease progression.

Statistical Analysis

Statistical analyses will be primarily descriptive in nature since thegoal of the study is to determine the MTD of Compound 1, or apharmaceutically acceptable salt thereof. Tabulations will be producedfor appropriate disposition, demographic, baseline, safety, PK, PD, andclinical activity parameters and will be presented by dose level andoverall. Categorical variables will be summarized by frequencydistributions (number and percentages of subjects) and continuousvariables will be summarized by descriptive statistics (mean, standarddeviation, median, minimum, and maximum).

Adverse events will be summarized by Medical Dictionary for RegulatoryActivities (MedDRA) system organ class and preferred term. Separatetabulations will be produced for all treatment-emergent AEs (TEAEs),treatment-related AEs (those considered by the Investigator as at leastpossibly drug related), SAEs, discontinuations due to AEs, and AEs of atleast Grade 3 severity. By-subject listings will be provided for deaths,SAEs, DLTs, and AEs leading to discontinuation of treatment.

Descriptive statistics will be provided for clinical laboratory, ECGinterval, LVEF, and vital signs data, presented as both actual valuesand changes from baseline relative to each on-study evaluation and tothe last evaluation on study. Shift analyses will be conducted forlaboratory parameters and ECOG PS.

Descriptive statistics will be used to summarize PK parameters for eachdose group and, where appropriate, for the entire population. Thepotential relationship between plasma levels of Compound 1, or apharmaceutically acceptable salt thereof and blood, plasma or urine 2HGlevels will be explored with descriptive and graphical methods.

Response to treatment as assessed by the site Investigators usingmodified IWG (for subjects with hematologic malignancies, such as MDS,MDS/MPN or AML). Two-sided 90% confidence intervals on the responserates will be calculated for each dose level and overall. Data will alsobe summarized by type of malignancy for subjects in the cohort expansionphase. Descriptive statistics will be used to summarize Ki67 levels fromtumor biopsies.

Study Results

Compound 1 had a cellular IC₅₀ value of 8-20 nM. Reduction in 2HG wasobserved following a single dose of Compound 1 in an IDH1 mutant R132Hxenograft model (FIG. 7A). In addition, Compound 1 reduced intracellular2HG in primary human IDH-mutated blast cells ex vivo (FIG. 7B).

TABLE 2 Cohort 1 Cohort 2 Cohort 3 Cohort 4 100 mg BID* 300 mg QD* 500mg QD 800 mg QD All patients (n = 4) (n = 4) (n = 4) (n = 5) (N = 17) Ontherapy, n 1 2 3 5 11  Discontinued, n 3 2 1 — 6 due to PD* 2 1 1 — 4due to AE*  1** — — —  1** due to — 1 — — 1 Investigator Decision Death≤28 days 3 2 1 — 6 after Compound 1 Discontinuation *AE, adverse event;PD, progressive disease; BID, twice daily; QD, once daily **AE =intracranial hemorrhage. This patient presented with right extremityweakness (upper and lower). The patient was hospitalized with a plateletcount of 11 on admission. The patient's status quickly deteriorated, aCT scan revealed left sided intraparenchymal hemorrhage. The patientdied 3 days later.

Patients in Table 2 have received median (range)=1.6 (0.4-5.7) of monthsof treatment.

TABLE 3 Demographic Characteristics of Patients of Table 2 All treatedpatients (N = 17) Age in years, median (range)  73 (42-87) Men/women, n8/9 Diagnosis, n R/R AML 17  ECOG performance status, n 0 6 1 9 2 2Number of prior 2 (1-5) chemotherapy regimens, median (range) Prior BMT,n 2 Abnormal cytogenetics, n 10 

One DLT (dose limiting toxicity) of grade 3QT prolongation at 800 mg QDwas observed. There were no associated cardiac symptoms, QTc returned tonormal following 3-day drug hold. The patient's dose was reduced to 500mg QD and remains on study with grade 1 QTc prolongation in completeremission (CR). Eight subjects experienced serious adverse events. At100 mg BID, 1 subject discontinued study due to an intracranial bleedattributed to disease progression and resulted in death. At 300 mg QD, 1subject experienced differentiation syndrome, recovered and was in CR.At 800 mg QD, 1 subject experienced tongue edema and QT prolongation(DLT described above), recovered and was in CR. All other SAEs relatedto disease progression resulted in death. For the patient thatexperienced differentiation syndrome, symptoms included fever anddyspnea. The patient was treated with steroids. Two events that led toPD are described as they are events that correlate to discontinuationsdue to AEs.

TABLE 4 Adverse Events Grade ≥3, All grades, AE n (%) n (%) At least 1adverse 11 (65) 14 (82) event Nausea 1 (6) 5 (29) Fatigue 1 (6) 5 (29)Dyspnea 2 (12) 5 (29) Vomiting 0 4 (24) Pyrexia 1 (6) 4 (24) Cough 0 4(24) Febrile 3 (18) 3 (18) neutropenia Diarrhea 0 3 (18)Electrocardiogram 1 (6) 3 (18) QT prolonged Notable grade ≥3 AEsinclude: hypotension 2 (12%), mental status changes 2 (12%), neutropenia2 (12%). AEs appear typical for this patient population. Other QTprolongations observed: Grade 1 prolonged QT in 100 mg cohort (thepatient had a history of right bundle branch block (R BBB) at studyentry; Grade 1 intermittent prolonged QT in 300 mg cohort; and Grade 3prolonged QT (DLT) in 800 mg cohort.

Compound 1 Exposure and 2HG Inhibition

FIGS. 8A and 8B shows the PK profile of Compound 1 following oraladministration. Compound 1 showed high plasma exposure, drugaccumulation and half-life of 182 hours. The plasma levels of 2HG werereduced to a normal range at all dose levels (up tp 98% inhibition). The2HG baseline was taken at Day −3 pre-treatment 2HG inhibition estimatedbased on 2HG pre-treatment level and AUC_(0-10 hr) post treatment. Forthe 100 mg BID and 300 mg QD Cohorts, 3 to 4 patients were measured pertime point and for the 500 mg QD Cohort, 1 to 3 patients were measuredper time point.

TABLE 5 Clinical Activity Assessed by Investigator Usine IWG AML and MDSCriteria Cohort 1 Cohort 2 Cohort 3 Cohort 4 100 mg BID 300 mg QD 500 mgQD 800 mg QD Total (n = 4) (n = 4) (n = 4) (n = 2) N = 14 CR 1 1 1 1 4Marrow CR — — 2 — 2 PR — — — 1 1 SD 3 2 1 — 6 PD — 1 — — 1 Overallresponse 1/4 1/4 3/4 2/2 7/14 rate (ORR) CR = complete response MarrowCR = ≤5% blasts in BM; no hematological recovery PR = partial responseSD = stable disease PD = progressive disease ORR = CR, Marrow CR and PR

Differentiation Effect in the Bone Marrow

FIGS. 9A-9C are images of aspirate from a 74 year old female patient whowas refractory to induction with 7+3. At baseline (FIG. 9A), her bonemarrow displayed monotonous cellularity, from the preponderance of blastcells. The inset shows the appearance of the blast cells on theaspirate. After 2 weeks of therapy (FIG. 9B), the core biopsy showedongoing hypercellularity, but clear evidence of maturation, asdetermined by the cells that have varied sizes and shapes, approximatingthe “field of flowers” appearance of a normal marrow. In the inset theaspirate no longer shows blast cells, but instead mostly myelocytes,which is evidence of differentiation. At this timepoint, given thereduction in blasts to <5% and preservation of neutrophils andplatelets, this patient met the criteria for a full CR. This wasmaintained at D28, which again showed hypercellularity, but withmaturation and no increase in blast cells (FIG. 9C).

While the foregoing invention has been described in some detail forpurposes of clarity and understanding, these particular embodiments areto be considered as illustrative and not restrictive. It will beappreciated by one skilled in the art from a reading of this disclosurethat various changes in form and detail can be made without departingfrom the true scope of the invention, which is to be defined by theappended claims rather than by the specific embodiments.

The patent and scientific literature referred to herein establishesknowledge that is available to those with skill in the art. Unlessotherwise defined, all technical and scientific terms used herein havethe same meaning as commonly understood by one of ordinary skill in theart to which this invention belongs. The issued patents, applications,and references that are cited herein are hereby incorporated byreference to the same extent as if each was specifically andindividually indicated to be incorporated by reference. In the case ofinconsistencies, the present disclosure, including definitions, willcontrol.

We claim:
 1. A method of treating advanced hematologic malignancies in asubject, each characterized by the presence of a mutant allele of IDH1,the method comprising administering to the subject in need thereof apharmaceutical composition comprising: (a) a compound(S)-N-((S)-1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide(Compound 1), or a pharmaceutically acceptable salt thereof, as part ofa solid dispersion; Form 1 of the Compound 1; or Form 2 of the Compound1; and optionally (b) one or more pharmaceutically acceptablecarrier(s).
 2. The method of claim 1, wherein the advanced hematologicmalignancies is selected from acute myelogenous leukemia (AML),myelodysplastic syndrome (MDS), myeloproliferative neoplasms (MPN),myeloproliferative neoplasms (MPN), chronic myelomonocytic leukemia(CMML), B-acute lymphoblastic leukemias (B-ALL), and lymphoma.
 3. Themethod of claim 1, wherein at least a particular percentage by weight ofCompound 1 is crystalline.
 4. The method of claim 3, wherein theparticular weight percentage of Compound 1 is 10%, 20%, 30%, 40%, 50%,60%, 70%, 75%, 80%, 85%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99%, 99.5%, or 99.9%.
 5. The method of claim 3, whereinthe particular weight percentage of Compound 1 is between 10% and 100%.6. The method of claim 1, wherein a particular percentage by weight ofCompound 1 is crystalline, and the remainder of Compound 1 is theamorphous form of Compound
 1. 7. The method of claim 1, wherein compound1 comprises a single crystalline form of Compound 1 or a mixture ofdifferent single crystalline forms.
 8. The method of claim 1, whereinCompound 1 is at least 90% by weight crystalline.
 9. The method of claim1, wherein Compound 1 is at least 95% by weight crystalline.
 10. Themethod of claim 1, wherein Compound 1 is at least 99% by weightcrystalline.
 11. The method of claim 1, wherein Form 1 of Compound 1 ischaracterized by the X-ray powder diffraction (XRPD) pattern shown inFIG. 1, and the data shown in Table
 1. 12. The method of claim 11,wherein the single crystalline form is characterized by one or more ofthe peaks shown in FIG. 1, and as shown in Table
 1. 13. The method ofclaim 11, wherein the single crystalline form is characterized by one ortwo or three or four or five or six or seven or eight or nine of thepeaks shown in Table
 1. 14. The method of claim 11, wherein Form 1 ischaracterized by the peaks identified at 20 angles of 8.6, 15.6, 18.5,20.6, 21.6, and 26.4°.
 15. The method of claim 11, wherein Form 1 ischaracterized by the peaks identified at 20 angles of 8.6, 15.6, 18.5,and 21.6°.
 16. The method of claim 1, wherein Form 2 of the Compound 1is characterized by the X-ray powder diffraction (XRPD) pattern shown inFIG. 4, and the data shown in Table
 2. 17. The method of claim 16,wherein Form 2 is characterized by one or more of the peaks shown inFIG. 4, and as shown in Table
 2. 18. The method of claim 16, whereinForm 2 is characterized by one or two or three or four or five or six orseven or eight or nine of the peaks shown in Table
 2. 19. The method ofclaim 16, wherein Form 2 is characterized by the peaks identified at 2θangles of 9.8, 11.6, 19.6, 22.5, 23.0, and 31.4°.
 20. The method ofclaim 11, wherein Form 2 is characterized by the peaks identified at 2θangles of 9.8, 11.6, 19.6, and 23.0°.
 21. The method of claim 1, whereinthe solid dispersion comprises a water-soluble polymer.
 22. The methodof claim 1, wherein the solid dispersion comprises one partiallywater-soluble polymer.
 23. The method of claim 21, wherein the polymeris a cellulose polymer.
 24. The method of claim 21, wherein the efficacyof treatment of advanced hematologic malignancies is monitored bymeasuring the levels of 2HG in the subject.
 25. The method of claim 1,wherein the subject is evaluated prior to and/or after treatment withthe pharmaceutical composition comprising: (a) Compound 1 or apharmaceutically acceptable salt thereof, as part of a solid dispersion;Form 1 of the Compound 1; or Form 2 of the Compound 1; and optionally(b) one or more pharmaceutically acceptable carrier(s), wherein themethod comprises determining the 2HG level in the subject.
 26. Themethod of claim 25, wherein the 2HG level is determined by spectroscopicanalysis.
 27. The method of claim 26, wherein the spectroscopic analysiscomprises magnetic resonance-based analysis.
 28. The method of claim 26,wherein the spectroscopic analysis comprises MRI and/or MRS measurement;sample analysis of bodily fluid; or by analysis of surgical material.29. The method of claim 28, wherein the bodily fluid comprises blood,plasma, urine, or spinal cord fluid.
 30. The method of claim 28, whereinthe surgical material is analyzed by mass-spectroscopy.
 31. The methodof claim 30, wherein the mass-spectroscopy comprises LC-MS or GC-MS. 32.The method of claim 1, wherein the advanced hematologic malignancies arecharacterized by a mutant allele of IDH1, wherein the IDH1 mutationresults in a new ability of the enzyme to catalyze the NAPH-dependentreduction of α-ketoglutarate to R(−)-2-hydroxyglutarate (2HG) in apatient.
 33. The method of claim 32, wherein the mutant IDH1 has anR132X mutation.
 34. The method of claim 33, wherein the R132X mutationis selected from R132H, R132C, R132L, R132V, R132S and R132G.
 35. Themethod of claim 33, wherein the R132X mutation is R132H or R132C. 36.The method of claim 1, wherein the advanced hematologic malignancies arecharacterized by a co-mutation selected from NPM1, FLT3, TET2, CEBPA,DNMT3A, and MLL.
 37. The method of claim 1, wherein the method comprisesadministering to the subject in need thereof a pharmaceuticalcomposition comprising Compound 1, or a pharmaceutically acceptable saltthereof, as part of a solid dispersion.
 38. The method of claim 1,wherein the method comprises administering to the subject in needthereof Form 1 of the Compound
 1. 39. The method of claim 1, wherein themethod comprises administering to the subject in need thereof Form 2 ofthe Compound 1.